Abstract: A BVD control programme based on the identification and removal of persistently infected (PI) animals is being undertaken in an area in the Rome province, where BVD outbreaks had been previously detected. It involves 174 mainly dairy herds, from which blood samples of all bovines older than 1 year are obtained through the national brucellosis and leukosis eradication programme. Samples sufficient to detect the presence of seropositive animals at a prevalence of 5% or more are initially screened for antibodies against BVD virus (BVDV) using an immunoenzymatic assay. Upon identification of seroreagents additional blood samples are tested from the 6-12-month age category not included in the initial samples. Animals are considered immunotolerant if BVDV is demonstrated twice at a minimum 30-day interval. When no seropositive animals are detected during the first serological screening the herd is declared BVD-free if a second testing, preferably carried on the same animals previously tested, confirms the seronegative status of the herd. At present 147 farms have been tested, of which 63 (42.9%) are negative with respect to antibodies against BVDV. Of the 84 remaining herds in which one or more seropositives are detected, 13 are classified as recently infected. In eight of these recently infected herds, 22 PI animals have been identified.

Abstract: Magnetic resonance imaging (MRI) is the most suitable modality for evaluation of infectious spondylitis. It is more sensitive than other imaging modalities for detecting presence and extent of such infections. Though it is not always possible to differentiate various infections on the basis of imaging findings alone, there are certain features which along with a good clinical background, can differentiate brucellar spondylitis from other spinal infections. It is useful to follow up such patients after specific chemotherapy to further confirm the diagnosis.

Abstract: Serum samples from 102 veterinarians and 191 pig farmers from the southern part of the Netherlands were investigated for antibodies against Brucella abortus, Leptospira spp, Streptococcus suis serotype II, Hantavirus (HV), and lymphocytic choriomeningitis virus (LCMV). All samples were collected in 1993 and stored until this study was performed. The prevalence of antibodies against B.abortus in veterinarians (4.5%) was significantly higher (P = 0.01) than in pig farmers (0%). None of the veterinarians (0%) and only one pig farmer (0.5%) had antibodies against Leptospira spp. Furthermore, significantly (P = 0.015) more veterinarians (6%) than pig farmers (1%) had antibody titres against muramidase-released protein (MRP),a protein of pathogenic S. suis serotype II strains. None of the veterinarians and a total of 3 (1.6%) pig farmers had antibody titres against HV. The prevalence of antibodies against LCMV tended to be higher in pig farmers (2.6%) than in veterinarians (0%) (P = 0.10). It can be concluded that the prevalence of antibodies against the investigated zoonotic agents in veterinarians and pig farmers is low.

Abstract: AIM: The susceptibility of common oral Candida species to saturated aqueous calcium hydroxide solution was studied. METHODOLOGY: The yeast species tested were C. albicans (16 strains). C. glabrata (three strains), C. guilliermondii (three strains), C. krusei (two strains), and C. tropicalis (two strains). At least one reference strain of each species was used; the others were clinical isolates either from persistent apical periodontitis or from marginal periodontitis. The susceptibility of Enterococcus faecalis (ATCC 29212) was studied for comparative purposes. Standardized inocula of the strains were incubated in aqueous calcium hydroxide solution, pH 12.4, for time-periods ranging from 5 min to 6 h. Volumes of 0.1 mL of the test suspension were cultured directly on Brucella blood agar and incubated in air at 37C. The plates were inspected for growth at 24 and 48 h and the colonies were counted. The time required to reduce the number of colony-forming units to less than 0.1% of the initial number was determined for each strain. RESULTS: The sensitivity of the C. albicans strains was generally low, with 16 h of incubation required to kill 99.9% of the colony-forming units. No differences in susceptibility between C. albicans strains isolated from root-canal infections and from periodontitis were found. Both strains of C. tropicalis were killed between 3 and 6 h of incubation, whilst strains of C. guilliermondii were killed after only 1020 min of incubation. All strains of C. glabrata survived 20 min, but not 1 h, of incubation, whilst 13 h were required to kill C. krusei. Compared with E. faecalis, all Candida spp. showed either equally high or higher resistance to aqueous calcium hydroxide. CONCLUSIONS: This study indicates that Candida spp. are resistant to calcium hydroxide in vitro, which may explain the isolation of yeasts from cases of persistent apical periodontitis.

Abstract: In this study, Brucella antibodies in bovine sera and milk were detected using the dot-immunobinding assay (DIA), the serum agglutination test (SAT), the Rose Bengal plate test (RBPT) and the milk ring test (MRT). For this purpose, a total of 116 paired blood and milk samples collected at the same time from 56 aborted and from 60 healthy dairy cows was examined. In DIA, a nitrocellulose membrane (NCM) was used as the solid phase. Antigen adsorbed on the NCM was extracted from Brucella abortus S99 by heat treatment. The results obtained by DIA were compared with those of SAT, RBPT and MRT. Of the 116 paired blood and milk samples, 24 were positive and 72 were negative by all tests used. Serum samples of six aborted cows were positive by DIA, SAT and RBPT but the milk samples were negative by DIA and MRT. Serum and milk samples of four aborted cows gave positive reaction only by DIA tests. The remaining six aborted cows were negative only by MRT and two of them were negative by both RBPT and MRT. Four sera of healthy cows were found to be positive only by SAT.

Abstract: Long term active surveillance of brucellosis was implemented in a rural area (Fokida) of Greece from 1989 to 1993 while the rural area of Grevena was selected as a control area. The computerised mapping programme was used to identify and protect the suspected animal brucellosis free zones. Health education of the inhabitants was further used to teach them how to avoid the risk factors. Three suspected brucellosis free zones were identified and two of them were successfully protected. The incidence for the 10 year period (1979-1988) was estimated at 1.4/1000/year for the study area and 1.6/1000/year for the control area. During the surveillance period the incidence in the study area dropped to 0.2/1000/year while in the control area it decreased to 1.0/1000/year. The methodology of identification and protection of suspected brucellosis free zones combined with health education proved to be efficient in reducing the incidence of the disease. The same methodology could be used in the country level, in countries where it is difficult to implement and maintain an animal control programme in the whole country.

Abstract: We investigated the farm factors associated with the prevalences of brucellosis and border disease (BD) in small-ruminant herds in the Madrid region of Spain. These infections were used as models of diseases of well-known and totally unknown distribution, respectively, to assess the association between the perception of the importance of a given disease on the relative contributions of veterinary services and the farmer's attitudes to its prevention. Sera, farming-management information and data concerning veterinary assistance and farmer characteristics were collected from 60 sheep or goat herds. The overall sero-prevalence of brucellosis was 5.7% (complement fixation) and for BD was 17.9% (ELISA test). The relationship between sero-positivity and the variables in the questionnaire was assessed by multivariable analysis using random-effects logistic-normal regression. 'Availability of veterinary services' was a major protective factor for brucellosis. In contrast, no association with veterinary services was observed for BD, whereas 'membership in a farmers' organization' (a variable associated with good farming practice and animal care) was a protective factor. 'Membership of a farmers' organisation' and two other farmer variables indicative of good husbandry ('youth' and 'schooling') were associated with a lower sero-prevalence of brucellosis in univariable analysis but they did not remain significant in the multivariable model. Our observations suggest that veterinary-activity variables predominate over non-specific protective farm factors related to good husbandry in the case the disease is subject to disease surveillance. This underscores the importance of organized control programs for veterinary services to be effective in terms of animal disease prevention.

Abstract: Different methods of extraction of bacterial DNA from bovine milk to improve the direct detection of Brucella by PCR were evaluated. We found that the use of a lysis buffer with high concentrations of Tris, EDTA, and NaCl, high concentrations of sodium dodecyl sulfate and proteinase K, and high temperatures of incubation was necessary for the efficient extraction of Brucella DNA. The limit of detection by PCR was 5 to 50 Brucella CFU/ml of milk.

Abstract: The deduced sequences of the Omp10, Omp16, and Omp19 outer membrane proteins of Brucella spp. contain a potential bacterial lipoprotein processing sequence. After extraction with Triton X-114, these three proteins partitioned into the detergent phase. Processing of the three proteins is inhibited by globomycin, a specific inhibitor of lipoprotein signal peptidase. The three proteins were radioimmunoprecipitated from [(3)H]palmitic acid-labeled Brucella abortus lysates with monoclonal antibodies. These results demonstrate that Omp10, Omp16, and Omp19 are lipoproteins.

Abstract: We present an extended analysis of the organization of green plant photosystem II and its associated light-harvesting antenna using electron microscopy and image analysis. The analysis is based on a large dataset of 16 600 projections of negatively stained PSII-LHCII supercomplexes and megacomplexes prepared by means of three different pretreatments. In addition to our previous work on this system [Boekema, E.J., van Roon, H., Calkoen, F., Bassi, R. and Dekker, J.P. (1999) Biochemistry 38, 2233-2239], the following results were obtained. The rotational orientation of trimeric LHCII at the S, M and L binding positions was determined. It was found that compared to the S trimer, the M and L trimers are rotationally shifted by about -20 degrees and -50 degrees, respectively. The number of projections with empty CP29, CP26 and CP24 binding sites was found to be about 0, 18 and 4%, respectively. We suggest that CP26 and CP24 are not required for the binding of trimeric LHCII at any of the three binding positions. A new type of megacomplex was observed with a characteristic windmill-like shape. This type III megacomplex consists of two C2S2 supercomplexes connected at their CP26 tips. Structural variation in the region of the central dimeric photosystem II complex was found to occur at one specific position near the periphery of the complex. We attribute this variation to the partial absence of an extrinsic polypeptide or one or more small intrinsic membrane proteins.

Abstract: Israel is one of the Mediterranean countries in which Brucellosis is endemic. As recently there has been a Brucellosis outbreak in a kibbutz, the aim of this study is to identify asymptomatic infected Kibbutz members, and to delineate the manner of infection in this setting. Therefore, all the asymptomatic Kibbutz members were screened by the Rose Bengal test for Brucellosis, while both patients and healthy members were asked to fill in a questionnaire in order to pinpoint the manner of infection, and signs and symptoms of the disease. In addition to the 14 patients with Brucellosis, 2 other Kibbutz members were also found to be infected by the screening tests. Analysis of the data of the questionnaires from 142 healthy and 16 patients disclosed that almost all of the infected patients (15/16) worked in the cowshed, as opposed to only 24 out of 142 (16.9%) of the healthy members. The infected tended to participate more in calf deliveries, and had contact with cow's blood and placenta, compared with the healthy subjects (P<0.001), while there were no significant differences with respect to having cuts on hands, or working in the cowshed without gloves. In addition, 15 out of 16 (93.8%) infected persons also drank unpasteurized milk, as compared with only 17 of the 142 (12%) healthy members (P<0.001), and thus were exposed to 2 major risk factors (working in the cowshed and consumption of unpasteurized milk). As the cows of the Kibbutz's cowshed were affected by Brucella melitensis (which usually affects flocks of goats and sheep rather than cows), the microbe was probably transmitted to the cowshed from neighboring flocks by wandering dogs, and then to the infected humans.

Abstract: We report an outbreak of brucellosis among 9 patients in a meat-packing plant. All patients developed chronic brucellosis characterized by severe chronic musculoskeletal pains, diffuse arthralgia, myalgia and recurrent bouts of fever, which we suggest may be related to a delay in antibiotic treatment. All the patients had a prolonged recovery requiring rehabilitation therapy.

Abstract: The epidemiology and control of Brucella melitensis in Malta was analysed using herd test data made available by the Veterinary Service of Malta. The eradication scheme commenced in 1987 with the introduction of a test and slaughter scheme using the Rose Bengal test. Herds registered with Malta Dairy Products Limited (MDP) showed a herd prevalence of 23% in 1987 which fell to less than 1.5% by 1993. Prevalence rose to 13% in 1995. Herds not delivering milk to the MDP showed an initial herd prevalence of 4% which fell below 1% in 1994, remaining under 2% in 1995. The epidemic in 1995 caused approximately 300 human brucellosis cases. Large herds and herds with small ruminants were most at risk to brucellosis infection. Seasonal fluctuation of prevalence was apparent. Increased enforcement of regulations and motivation of farmers would accelerate eradication of brucellosis in Malta.

Abstract: In view of treating intracellular Brucella infections, microspheres made of poly(lactide) (PLA) and poly(lactide-co-glycolide) (PLGA) were developed as delivery system for the cationic and highly hydrophilic antibiotic gentamicin sulphate. Drug microencapsulation by spray drying yielded microspheres with regular morphology, an average particle size of approximately 3 micrometer and encapsulation efficiencies of up to 45%. Different copolymers of similar molecular weights gave varying encapsulation efficiencies and particle size distributions. The encapsulation efficiency generally increased with polymer hydrophilicity, except for the hydrophilic copolymer PLGA50:50H carrying carboxylic end groups. Encapsulation also depended on the pH value of the aqueous drug solution to be encapsulated. Moreover, increasing nominal gentamicin sulphate loading yielded lower efficiencies. For comparison, some formulations were also prepared by a (W(1)/O)W(2)-solvent evaporation method, which yielded lower encapsulation efficiencies, in the order of 13%. Finally, drug bioactivity was found to remain intact after microencapsulation, MS storage and MS incubation in aqueous medium. The results suggest that PLA/PLGA microspheres prepared by spray drying may be an appropriate delivery system for gentamicin sulphate to be used in the treatment of intracellular Brucella infections.

Palanduz A, Palanduz S, Guler K, Guler N. Brucellosis in a mother and her young infant: probable transmission by breast milk. International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases. 2000; 4(1); 55-6. [PubMed: 10689217].

Abstract: Brucellosis, although primarily a zoonotic infection, is also a threat for human health. Infection can be transmitted to humans through direct contact with infected animals, products of conception, or animal discharges, and through consumption of potentially infected milk, milk products, or meat. Human-to-human transmission is rare. There have been case reports of transmission via blood transfusion and bone marrow transplantation from infected donors. Sexual intercourse is a possible means of transmission. Neonatal infection can be acquired transplacentally or during delivery. This report describes a mother with brucellosis who probably transmitted the infection to her 3-month-old baby by breast milk.

Wyatt HV. Royal Navy surgeons and the transmission of brucellosis by goats' milk. Journal of the Royal Naval Medical Service. 1999; 85(2); 112-7. [PubMed: 10707453].

Abstract: In 1997, 7 cases of laboratory-acquired Brucella melitensis infections were detected among the hospital personnel of a medical centre serving an endemic area in southern Israel. Although the onset of symptoms in 6 of the 7 patients occurred during a 2-week period, suggesting a point source exposure, biotype analysis showed that the outbreak was caused by 3 different B. melitensis serovars, indicating multiple exposures. Review of the laboratory records showed that during 1997, the microorganism was recovered from 146 blood and synovial fluid cultures, and that during the 2 months in which the laboratory-acquired cases occurred (April and June), 53 of 530 positive aerobic blood culture bottles (10.0%) grew B. melitensis. The epidemiological investigation did not reveal the source of the outbreak, and no noticeable breaches in laboratory safety practices could be demonstrated. It is concluded that in areas endemic for brucellosis, hospital personnel are frequently exposed to Brucella microorganisms. Under these circumstances, significant morbidity may occur despite observance of recommended safety practices. Biotyping of Brucella isolates may contribute to the elucidation of complex epidemiological situations.

Abstract: The objective of this study was to determine if B. abortus rough mutant strains RB51 and rfbK are eliminated in goat milk. Thirty milk goats were divided into two groups. Group I was inoculated with 4x10(10) cfu/ml of B.abortus RB51 strain and Group II with 1x10(9) cfu/ml of B. abortus rfbK strain by subcutaneous route in the right axilary region. Milk samples were taken aseptically on a daily basis for the first 30 days and weekly for the second and third months. The samples were inoculated on Farrell's media and incubated under 5-10% CO(2) at 37 degrees C for 10 days. The suspicious colonies were recultured in Farrell's media. There were no isolations of bacteria with characteristics of Brucella from any of the milk samples collected during 90 days of the study. It was concluded that neither of the strains used at these doses were eliminated by milk in goats inoculated during lactation.

Abstract: A modified procedure is described for performing the MRPAH (mixed reverse passive antiglobulin haemagglutination) reaction as a simple micro-method to measure the classes of bacterial antibodies. This 'bacterial dilution procedure' gave results closely correlated with those obtained by the 'serum (sample) dilution procedure' previously reported and with great economy of materials, labour and time. The method was used to investigate human serum antibodies to Br. abortus and S. enteritidis and serum and secretory antibodies to Strep. mutans. The good reproducibility of the MRPAH reaction was demonstrated by re-examining brucellosis sera tested one year previously. MRPAH was sufficiently sensitive to demonstrate the small amounts of IgG and IgM antibodies to Strep. mutans in human colostrum and early milk. A rise of antibody levels in the different immunoglobulin classes G, A and M was readily demonstrated in sera from individuals with salmonellosis.

Abstract: Thirteen cows, which had been vaccinated as calves with strain 19, were revaccinated twice or three times as adults with I x 109 cfu of B. abortus strain RB51. Their serological responses following adult vaccination remained negative to conventional brucellosis surveillance tests. Vaccination with strain RB51 during the eighth month of pregnancy did not induce abortion, although strain RB51 was recovered from milk for up to 69 days after vaccination. In a parallel study, thirteen 8- to 10-month-old heifers were vaccinated as calves with 109 cfu of strain RB51. The heifers remained seronegative to conventional brucellosis surveillance tests but antibody responses to RB51 could be demonstrated using an indirect ELISA. This study showed that multiple vaccination with strain RB51 did not induce seroconversion to brucellosis surveillance tests. In addition, this study suggests that 109 cfu of strain RB51 is safe for use in pregnant cattle.

Abstract: The polymerase chain reaction (PCR) was used to diagnose goat brucellosis and compare its sensitivity against some of the most commonly used serological and bacteriological techniques. Twenty two female and one male out of 300 clinically healthy, mixed-breed goats were randomly chosen from a ranch located at Marín, Nuevo León, Mexico. Milk and blood samples were taken from each animal and used to obtain both microbiological cultures and DNA of the pathogen, and sera was tested against Rose Bengal antigen (RBT). Results showed that 86% of the blood samples were positive on the PCR test, while 60% were positive on the serological test. The pathogen was isolated from only one blood culture. Sixty four percent of the milk samples were positive on PCR tests, but failed to yield bacteria in culture. Biochemical and PCR specific assay demonstrated that Brucella abortus biovar 1 was associated with the infection. This study demonstrates the higher sensitivity of PCR over RBT and blood culture and its potential towards a rapid identification of Brucella strains.

Abstract: A multiplex amplification and detection platform for the diagnosis of Mycobacterium bovis and Brucella abortus infection simultaneously in bovine milk and nasal secretions was developed. This system (designated the bovine pathogen detection assay [BPDA]-PCR) consists of duplex amplification of species-specific targets (a region of the BCSP31K gene of B. abortus and a repeat-sequence region in the hsp65 gene of M. bovis, respectively). This is followed by a solid-phase probe capture hybridization of amplicons for detection. On the basis of spiking experiments with normal milk, the analytical sensitivity of the assay was 800 CFU equivalents/ml of milk for B. abortus and as low as 4 CFU equivalents per ml of milk for M. bovis. BPDA-PCR was validated with 45 liver samples from lemmings experimentally infected with B. abortus. The assay sensitivity, based on culture status as a "gold standard," was 93.9%. In this experiment, BPDA-PCR also identified five culture-negative liver samples as positive (41.7%). Field studies for the evaluation of BPDA-PCR were performed with samples from dairy animals from geographically distinct regions (India, Mexico, and Argentina). A high prevalence of shedding of B. abortus (samples from India) and M. bovis (samples from Mexico) was identified by BPDA-PCR. In samples from India, B. abortus shedding was identified in 86% of milk ring test-positive animals (n = 15) and 80% of milk ring test-negative cows (n = 5). In samples from Mexico, M. bovis was identified by PCR in 32.6% of pools (n = 46) of milk that each contained milk from 10 animals and in 56.2% of nasal swabs (n = 121) from cattle from tuberculin test-positive herds. In contrast, the Argentine cattle (n = 70) had a modest prevalence of M. bovis shedding in nasal swabs (2.9%) and milk (1.4%) and of B. abortus in milk (11.4%). On the basis of these analyses, we identify BPDA-PCR as an optimal tool for both screening of herds and testing of individual animals in a disease eradication program. A combination of the duplex assay, screening of milk samples in pools, and the proposed algorithm provides a highly sensitive, cost-effective, and economically viable alternative to serological testing.

Abstract: Four seed lots and fourteen batches of Brucella melitensis Rev 1 and B. abortus B19 living anti-Brucella commercial vaccines obtained from six Spanish laboratories were tested in vitro and in vivo in the reference mouse model for quality control. All the strains tested showed the characteristic morphology of their respective Rev 1 or B19 reference strains with the exception of three Rev 1 strains: seed lot SL2 and commercial vaccine R3, in which giant colonies were predominant, and commercial vaccine R5, in which 5% rough colonies were detected. Strains SL2 and R5 (but not the R3) had a deficient activity when tested in the mouse model. All strains but two (Rev 1 strain R1 and B19 strain B2) had standard resistance/ sensitivity patterns to streptomycin and penicillin G. Strains R1 and B2 had an increased resistance to penicillin when incubated in a 10% CO2 atmosphere and both strains showed an increased residual virulence in mice. As residual virulence and immunogenicity in mice were not always correlated one another nor with the in vitro tests, all tests should be performed to control properly the anti-Brucella live vaccines. A computerized statistical procedure to calculate the residual virulence of vaccines is proposed as an alternative to that used in the current method.

Abstract: A study between August 1995 and December 1997 included 343 dairy cattle on 20 farms in the Dar es Salaam region and 2289 zebu cattle on 39 bomas in the Lugoba area (coast region). The aim was to establish the prevalence of bovine tuberculosis (Mycobacterium bovis) and bovine brucellosis (Brucella abortus). In the single intradermal tuberculin test (SIT), 0.9% (3/343) of the animals in Dar es Salaam tested positive and 1.2% (4/343) were doubtful. Positive reactors were found in 10% (2/20) of the farms. In the Lugoba area, 0.6% (14/2206) were positive and 6.8% (149/2206) doubtful, positive cases being found in 21% (8/39) of all bomas. In the slow agglutination test (SAT) for B. abortus, 14.1% (48/341) of the serum samples reacted positively in Dar es Salaam and 2.3% (8/341) were doubtful. Positive SAT reactors were identified on 25% (5/20) of the dairy cattle farms. In the Lugoba area, 12.3% (273/2221) proved to be positive SAT reactors and doubtful reactions were observed in 2.9% (64/2221). SAT-positive animals were detected on 87% (34/39) of all bomas. The prevalence in single herds in Dar es Salaam varied from 4.3% to 5.3% for the SIT and from 2.2% to 50% for the SAT. The prevalence in single herds in Lugoba area was between 1.1% and 2.9% for SIT and from 1.4% up to 62.1% for SAT. The two cattle populations differed significantly (p < 0.001) in the prevalence of both bovine tuberculosis and bovine brucellosis. Two cows that were positive reactors were slaughtered and subjected to post-mortem examination, and organ samples were bacteriologically cultured. The occurrence of Mycobacterium tuberculosis was confirmed by polymerase chain reaction (PCR) in both cows.

Abstract: OBJECTIVE: To evaluate biosecurity practices of cow-calf producers. DESIGN: Cross-sectional survey. SAMPLE POPULATION: 2,713 cow-calf operations were used in phase 1 of the study, and 1,190 cow-calf operations were used in phase 2. PROCEDURE: Producers were contacted for a personal interview between Dec 30, 1996 and Feb 3, 1997 regarding their management practices. Noninstitutional operations with 1 or more beef cows were eligible to participate in the study. Producers who participated in the first phase of the study and who had > or = 5 beef cows were requested to continue in the study and were contacted by a veterinarian or animal health technician who administered further questionnaires. All contacts for the second phase of the study were made between Mar 3, 1997 and Apr 30, 1997. Additional data on use of various vaccines, testing of imported cattle for brucellosis, Mycobacterium paratuberculosis, bovine viral diarrhea, and tuberculosis as well as potential for feed contamination were collected during the second phase of the study. RESULTS: Producers commonly engaged in management practices that increased risk of introducing disease to their cattle such as importing cattle, failing to quarantine imported cattle, and communal grazing. Producers inconsistently adjusted for the increased risk of their management practices by increasing the types of vaccines given, increasing the quarantine time or proportion of imported animals quarantined, or increasing testing for various diseases in imported animals. CONCLUSIONS AND CLINICAL RELEVANCE: Cow-calf herds are at risk for disease exposure from outside sources when cattle are introduced to the herd, and producers do not always adjust management practices such as vaccination schedules and quarantine procedures appropriately to minimize this risk. Veterinary involvement in education of producers regarding biosecurity risks and development of rational and economical biosecurity plans is needed.

Abstract: Zoonotic diseases constitute a public health problem throughout the world, particularly in the tropics, where their control is restricted by inadequate infrastructure and financial resources. Additionally, there is a lack of information on their significance and distribution. This study, conducted jointly by the Ministries of Health and Animal Resources, aimed to assess the prevalence of zoonotic diseases in Burkina Faso. The data were taken from internal reports of each ministry covering the period January 1-December 31 1996 for the Ministry of Health and for January 1-December 31 1997 for the Ministry of Animal Resources. Zoonotic diseases were divided into viral (rabies, yellow fever, HIV infection/AIDS, and measles), bacterial (tuberculosis, brucellosis, and anthrax) and parasitic (cysticercosis, toxoplasmosis, and leishmaniasis). For the period under study, the following diseases were reported by the Ministry of Health, tuberculosis, 1314 cases; anthrax, 145 cases; leishmania, 271 cases; rabies, 110 cases; and measles, 46490 cases. The Ministry of Animal Resources reported 69% of rabies cases occurred in dogs; cysticercosis occurred in swine at a prevalence of 0.57%; the prevalence of tuberculosis in cattle, small ruminants and pigs was 0. 13, 0.013, and 0.029%, respectively; the prevalence of anthrax and echinococcosis was 0.012 and 0.007%, respectively; and finally, the prevalence of bovine brucellosis was 8% in the peri-urban areas. This study revealed that there was a lack of collaboration between the organisational structures and workers in both ministries involved in the control of zoonoses. Links between the two ministries in the field of public health need strengthening.

Abstract: Usually symptoms of brucellosis are nonspecific and characterized by a wide range of complaints. Although the disease in Israel is almost exclusively food borne (caused by Brucella melitensis in unpasteurized goat milk products) so the main route of infection is the gastrointestinal tract, but gastrointestinal complications are rare, and only sporadic cases of ileitis or colitis have been described. We present a 43-year-old woman with an acute abdomen, probably due to diverticulitis. It was diagnosed only after blood cultures were positive for Brucella melitensis. We believe that its protean manifestations should be considered in addition to the other bizarre presentations of this disease, important in our region.

Abstract: Retrospectively we evaluated the records of 68 children with brucellosis. We found 58.2% had consumed unpasteurized milk and dairy products. Nonspecific manifestations included: arthralgia (78%), fever (75%) and sweating (60%). Localized manifestations included limping (75%) and arthritis (54%). Leukopenia was found in 51% of children and anaemia in 24%. Brucella species was cultured for blood of 16 (23.5%) patients. Combination therapy containing streptomycin was more effective than gentamicin combinations.

Abstract: Fluorescence polarization immunoassay (FPA) is a homogeneous immunoassay useful for rapid and accurate detection of antibody or antigen. The principle of the assay is that a fluorescent dye (attached to an antigen or an antibody fragment) can be excited by plane-polarized light at the appropriate wavelength. As a rule, a small molecule rotates faster when in solution than a larger molecule. The rotation rate may be assessed by measuring light intensity in the vertical and horizontal planes. Generally, the time it takes a molecule to rotate through a given angle is an indication of its size. When a small molecule that rotates rapidly is bound to a larger molecule, the rotation rate is decreased and this decrease is measured. Because it is a primary antigen-antibody interaction, the rate of reaction is very rapid and usually a result may be obtained in minutes. This technology was applied to the detection of antibody to Brucella abortus in serum and milk, providing for the first time a rapid primary binding assay that is cost effective for use in the field.

Abstract: Despite severe and persistent criticism of her research, Alice Evans persevered in her pioneering work on the bacterial contamination of milk, identifying the organism that caused undulant fever and demonstrating that drinking unpasteurized cow's milk could transmit the disease, undulant fever, to humans. The opprobrium that Alice Evans endured was unrelenting, even after her election as the first President of the Society of American Bacteriologists, (now the American Society for Microbiology), but she remained undeterred, a true heroine of American microbiology and a magnificent public health worker.

Abstract: In Sicily, as in other Mediterranean areas, livestock represents one of the most important resources for the island economy. This sector involves more than 16,000 farms of cattle and 10,000 farms of sheep and goats (respectively 6% and 15% of national production) which are actually increasing their number. Most livestock in Sicily is owned by small holders and pastoralists. Regional production of milk feeds some industries, which involve a large and increasing occupational area. Due to its peculiar geographic aspect Sicily is constituted by hill areas with sporadic grasses, therefore extensive grazing methods represent an ancient, traditional practice for using poor lands. For the control of infectious diseases Veterinary Services (VS) are based on the public regional network that is coordinated by the Ministry of Health in Rome. Even if Sicilian VS had to solve many constraints related to traditional "pastoral management system", to the lack of any sanitary background of this sector, to unknown data on the numbers of the whole livestock, the effort done in the last years has allowed to monitor for brucellosis, bovine leukemia virus (BLV), bovine TBC, swine vescicular disease (MVS) and other diseases for the majority of the farms. Tick-borne diseases (TBDs) still represent a serious impediment to the improvement of meat and milk production in the region. Every year, several outbreaks related to Theileria, Babesia or Anaplasma infections in cattle are recorded. Imported breeds pay the most expensive tribute often with a mortality rate of 100%. In the last five years more than 170 outbreaks of TBDs have been notified by our Institute although the pathogen prevalence and economical impact in the Sicilian livestock is still unknown. The outdoor grazing of the animals, far away from observation, a subclinical course of the disease, can in fact create difficulties in discovering infection and therefore the therapy is often too late. New diagnostic and control methods (PCR, vaccination) are being developed at the Istituto Zooprofilattico Sperimentale of Sicily in cooperation with other national and international laboratories (such as the University of Utrecht).

Abstract: Brucellosis is known to occur in Yemen but its epidemiology has not been extensively studied. The present investigation examined risk factors for human brucellosis in Yemen using a hospital-based case-control study. A total of 235 consecutive patients with brucellosis attending the Central Health Laboratory in Sana'a, Yemen, were matched in respect of age, sex, and place of residence, rural or urban, with 234 controls selected from individuals attending the Central Health Laboratory for unrelated health problems. Clinical information on patients and controls was supplemented with occupational and socio-economic data obtained by interview of cases and controls using a standard questionnaire. After controlling for confounding factors significant risk factors for infection related to occupation as a farmer (OR 2.5 (95% CI 1.4-4.5, P < 0.0001)), shepherd (OR 7.8 (95% CI 1.0-61, P 0.05)) or microbiologist (OR 24.5 (95% CI 2.9-204, P 0.003)); and drinking fresh milk (OR 2.0 (95% CI 1.3-4.3, P 0.001)) and laban (OR 22.7 (95% CI 1.7-4.2 P < 0.0001)). Taking other milk products and offal were not risk factors. Socio-economic and educational factors were also independent risk factors. Occupational, food and socio-economic risk factors significantly confounded one another. Yemen shares some but not all of the risk factors of neighbouring countries. The interrelation between the various factors is complex and studying any one in isolation may give a false impression of its public health significance. Control through education of the population to minimize exposure to, and contact with, animals and their milk and milk products and to boil milk before drinking it or using it to make buttermilk, would be difficult as these would represent such fundamental changes to established patterns of behaviour of this society. Ideally there would be a campaign to control the infection by animal vaccination but the costs and logistic difficulty would be great. Presently there is a clear need for doctors in Yemen to be made aware of the frequency of this infection, the means available for clinical and laboratory diagnosis and effective treatment, while strategies to control the disease in Yemen are formulated and field tested.

Abstract: Samples from 2427 cattle, 661 goats, 104 sheep, 98 camels and 82 horses were screened for brucella infections by the Rose Bengal Test and positive reactors confirmed by the complement fixation test. In cattle, the highest individual seroprevalence was in dairy herds kept under the intensive husbandry system, with an individual prevalence of 8.2% and unit (herd) seroprevalence of 35.9%. This was followed by the pastoral husbandry system in the Western Lowlands with 5.0% individual but a higher unit (vaccination site) prevalence of 46.1%. The lowest was in the mixed crop-livestock system in the Southern Highlands with individual 0.3% and unit (village) prevalence of 2.4%. In sheep and goats, no positive animals were detected in the mixed crop-livestock areas. In the Eastern Lowlands individual prevalences of 3.8% (goats) and 1.4% (sheep) and unit prevalence of 33.3% (goats) and 16.7% were found, while 14.3% of individual goats and 56.3% of the units in the Western Lowlands were positive. No positive horses were found. The present study documents the first serological evidence of Brucella spp. infection in camels (3.1%) in Eritrea.

Abstract: Multilocus enzyme electrophoresis (MLEE) of 99 Brucella isolates, including the type strains from all recognized species, revealed a very limited genetic diversity and supports the proposal of a monospecific genus. In MLEE-derived dendrograms, Brucella abortus and a marine Brucella sp. grouped into a single electrophoretic type related to Brucella neotomae and Brucella ovis. Brucella suis and Brucella canis formed another cluster linked to Brucella melitensis and related to Rhizobium tropici. The Brucella strains tested that were representatives of the six electrophoretic types had the same rRNA gene restriction fragment length polymorphism patterns and identical ribotypes. All 99 isolates had similar chromosome profiles as revealed by the Eckhardt procedure.

Abstract: The risk of zoonotic disease transmission when handling livestock or animal products is substantial. In industrialized countries, the classical zoonotic diseases such as tuberculosis or brucellosis are no longer in the foreground. Latent zoonoses such as salmonellosis and campylobacteriosis can cause serious disease in humans and have become a major public health problem during the past years. Since animals infected with these pathogens show only mild transient disease or no clinical signs at all, new concepts in the entire production line ("stable to table") are necessary in order to avoid human infection. Two emerging viruses with zoonotic potential--avian influenza virus and Nipah virus--have been found in Asia in 1997 and 1999. Both diseases had a major impact on disease control and public health in the countries of origin. In order to cope threats from infectious diseases, in particular those of public health relevance, a combined effort among all institutions involved will be necessary. The proposed "European Center for Infectious Diseases" and the "Swiss center for zoonotic diseases" could be a potential approach in order to achieve this goal.

Cheville NF. Development, testing and commercialization of a new brucellosis vaccine for cattle. Annals of the New York Academy of Sciences. 2000; 916; 147-53. [PubMed: 11193615].

Abstract: Vaccines used against brucellosis do not generally protect completely against infection or abortion. Genetic analysis has revealed differences in arrangements of DNA sequences between these vaccine strains and the virulent parent strain and permits the specific identification of field isolates of B. abortus as wild-type or vaccine strain. B. abortus strain 19 is a low-virulence, live vaccine developed for use in cattle. Although it is effective, strain 19 vaccine had a tropism for the placenta and caused abortion when given to pregnant cows, was infectious for humans, and caused serologic responses in calves that could not be differentiated from those in cattle infected with natural field strains. In the mid-1980s the need for a new vaccine emerged when the USDA increased its efforts in brucellosis eradication. In the 1990s, research on biosafety, vaccine efficacy and field application rapidly established the fact that strain RB51 is protective in cattle at doses comparable to those of strain 19. Thus, Brucella abortus strain RB51 is the vaccine of choice against brucellosis of cattle in the United States. Studies have established the relative efficacy of strain RB51 vaccine on bison, and the vaccine has also been accepted for use in commercial bison herds in the U.S.

Abstract: A case of severe chronic spondylitis of the lumbar spine caused by Brucella abortus is presented. Infection was probably associated with ingestion of raw cow's milk. A technetium bone scan provided evidence of activity as early as two years before the radiographic abnormalities were evident. Although rarely seen in brucellosis, a large left psoas abscess formed. After extensive resection and debridement of all necrotic tissue, along with prolonged use of oral doxycycline, the patient recovered completely. The clinical, laboratory, therapeutic, and pathogenic aspects of Brucella spondylitis are discussed.

Abstract: A polymerase chain reaction test was developed to detect Brucella spp. directly in milk and cheese and optimized using primers for the BSCP-31 gene. A total of 46 cheese samples produced with sheep and goats milk were assayed, and Brucella spp. was detected in 46% of them, especially in cheese made from sheep milk. This method is of remarkable epidemiologic interest because it is an indirect test indicating the sanitary quality of milk used in dairy industries. The method showed good sensitivity and specificity. It is faster and less expensive than the conventional bacteriological assays.

Abstract: The half-life of bovine immunoglobulins was established in a group of 60 calves suckled on non-infected dams immunised with antigens to Brucella abortus. The half-life of the "incomplete antibody" as determined by the anti-bovine globulin test was 20.9 days. Up to 45 days, the mean half-life of antibodies in the calves was 17.1 days; from 46 to 99 days it was 21.1 days and for periods exceeding 100 days it was 24.4 days. In 39 calves born of and suckled on infected dams the mean half-life of antibodies was 22.4 days. Up to 100 days the figure was 19.5 days; over 100 days it was 22.8 days. There was no significant difference in the mean half-life of antibody between calves from infected and non-infected dams. In both cases the antibodies appear to be catabolised at a progressively slower rate.

Abstract: To determine the sanitary risk to human health, 59 sera samples of clandestine slaughtered porks were examined through serologic procedures and have demonstrated to have anti-Brucella antibodies and antibodies titles suggestive of brucellosis infection. Surveillance measurements are recommended to prevent potential risk of zoonotic infection.

Abstract: A fluorescence polarization assay (FPA) was used to test whole blood samples prepared by mixing blood cells from cattle without exposure to Brucella abortus (B. abortus) with sera from animals with confirmed (bacteriologically) infection. A cut-off value between negative and positive values was initially established to be 87.2mP. This value was changed to 95mP to increase assay specificity without loss of sensitivity when testing blood samples from negative animals. The FPA technology was applied to whole blood samples in the field and to stored whole blood samples using two diluent buffers. Relative sensitivity and specificity values for the FPA performed in the field, based on buffered antigen plate agglutination test and competitive enzyme immunoassay results were 95.3 and 97.3%, respectively. However, to obtain maximum sensitivity and specificity, a cut-off value of 105mP was determined for fresh whole blood samples. The relative sensitivity and specificity values of the FPA when testing stored whole blood samples were 100% each using a 95mP cut-off.The usefulness of the FPA for testing whole blood samples in the field was demonstrated.

Abstract: The fact that non-specific milk ring tests were occasionally observed in tank milk provided the reason for doing a survey to detect the possible cause. Among others, non-specific reactions were found to occur after treatment with Estrumate. A regressing corpus luteum may also give rise to non-specific milk ring tests in animals which have not been treated with Estrumate.

Abstract: BACKGROUND: The onset of a large number of cases of brucellosis among employees at a slaughterhouse in Zaragoza where sheep from livestock culling procedures were being slaughtered, led to an investigation being made for the purpose of shedding light on the cause of this outbreak. METHODS: This study was organized into three parts: 1) Description of the outbreak 2) Study of the slaughterhouse layout and activity as regards the number of animals slaughtered, the employee workload and the degree of risk involved in each working area and 3) Unmatched case-control study. RESULTS: No significant differences were found among the attack rates by the sections of the slaughterhouse where the employees in question worked. The slaughtering analysis revealed a concomitant fluctuation between the slaughter of culled sheep and the epidemic. The case-control study revealed no significant differences for the risk factors involving working in a risk area, the use of protective measures and cuts/wounds. CONCLUSIONS: This outbreak is related to the slaughter of culled sheep. The fact that no significant difference were found by sections or due to working in a risk area is compatible with an overall explanation. Nor were any differences among the employees found to exist with regard to cuts/wounds or the use of protective measures, which focuses importance on airborne spread.

Abstract: Sera from 58 odontocetes taken in fisheries off Peru in 1993 to 1995 and from 24 cetaceans stranded along the Spanish coast of the Mediterranean in 1997 to 1999 were tested for the presence of Brucella species antibodies in competitive and indirect ELISAS (cELISA and iELISA). Among the animals from Peru, 21 of 27 (77.8 per cent) Lagenorhynchus obscurus, three of six Delphinus capensis, one of two inshore and two of three offshore Tursiops truncatus and five of 20 (25 per cent) Phocoena spinipinnis were positive in the cELISA. Brucella species antibodies were also observed in two of 16 (12.5 per cent) Stenella coeruleoalba and in one of two Ttruncatus from the Mediterranean. These data provide the first evidence for the presence of cetacean brucellae in the south Pacific Ocean and the Mediterranean Sea.

Abstract: A total of 114 beef and baby beef samples were examined. The samples included ground baby beef, mixed ground baby beef and pork, and chopped and shaped meat. The samples were analyzed from 30 different grocery stores in Zagreb, Croatia. The object of this study was to evaluate the prevalence of Escherichia coli O157:H7 in the samples that can enhance the potential risk of outbreaks of hemorrhagic colitis and hemolytic uremic syndrome. The results in all tested samples of E. coli O157:H7 were negative. A single sample was positive in a latex agglutination test using antiserum to O157:H7. It was identified as Proteus vulgaris at the Pasteur Institute, Paris, France. This result correlates positively with cross-contamination with Yersinia enterocolitica 09, Brucella abortus, Salmonella type N, and Pseudomonas maltophila.

Abstract: An indirect enzyme-linked immunosorbent assay (ELISA) for the detection of Brucella abortus antibodies in bovine bulk milk samples was evaluated. About 31 individual milk samples from B. abortus infected cows were diluted into bulk milk from a brucellosis free herd. Individual milk samples obtained from 96 negative or positive herds to ELISA or Brucella ring test (BRT), were tested by ELISA. All positive cows were bled and serum samples were tested by the complement-fixation test (CFT) which was considered the definitive test. A herd was considered infected if at least, one cow was positive in the CFT. Four samples were negative in the BRT at the dilution 1:10 but positive in the ELISA. For samples positive in both tests, BRT titers ranged from 1:10 to 1:480 while ELISA titers ranged from 1:10 to 1:3200.Using bulk milk samples, the sensitivity of the ELISA (98.1%) was higher than the BRT (72.2%) but the specificity of BRT (90.5%) was not statistically different (P=1.0) from the ELISA (88.1%). The implications of the results for brucellosis control are discussed.

Abstract: Brucellosis is a zoonotic disease of worldwide distribution. Despite its control in many developed countries the disease remains endemic in Saudi Arabia where the national seroprevalence of the disease is 15%. In Saudi Arabia the disease is introduced through uncontrolled importation of animals that are poorly screened for the disease. Every year the Kingdom imports a few million heads of sheep and goats for sacrifice during Hajj from Africa, India, and Autstralia. Brucella melitensis remains the principle cause of human brucellosis in Saudi Arabia, causing 88-93% of the cases. Recent national statistics indicate that the disease incidence in humans is close to 40 cases per 100,000. The eradication of human brucellosis in Saudi Arabia will ultimately depend on the eradication of animal brucellosis. There is an urgent need for a national program for controlling brucellosis in the Kingdom. The components of this program will include recruitment and training of qualified veterinarians, development of an adequate number of animal quarantine centers and implementing legislation to control marketing and movement of animals.

Abstract: OBJECTIVES: To evaluate BACTEC 9050 system capacity for detection of bacteremia due to Brucella spp. MATERIAL AND METHODS: 16 febrile patients were studied during an epidemic infection for Brucella spp. Suspicious of brucellosis was serologically confirmed with Rose Bengal test and agglutination tube test.Only one blood culture was processed per patient. Ten millilitres of blood were inoculated in a Bactec Plus aerobic/F bottle and incubated in BACTEC 9050 automatic system. The bottles were kept in incubation during 21 days, and they were subcultured when the machine detected its growth; if not, a blind subculture was performed after 21 days. RESULTS: 13 of 16 patients showed bacteriemia (81.2%): 11 patients were detected by BACTEC 9050 system and 2 patients by blind subculture after 21 days. A positive result appeared in 149.8 hours (6.2 days) as a mean. Earlier detections were seen in 83 hours (3.4 days) and the latest ones at 245 hours (10.2 days). The aetiology agent of the epidemic infection was Brucella mellitensis biotype 1.We found significative differences comparing the BACTEC 9050 with BACTEC 9120/9240 systems (p<0.03). CONCLUSIONS: Incubation protocols of 5 days are not useful for BACTEC 9050 system in the case brucellosis suspicious. Protocols of 7 would detect only 69.2% of the cases.

Abstract: A hot saline antigenic extract (HS) from Brucella ovis was encapsulated in poly-epsilon-caprolactone microparticles (PEC), and tested as a vaccine against B. ovis and B. abortus infections in mice. Subcutaneous but not oral administration in BALB/c mice of the HS-PEC induced high amounts of IFN-gamma and IL-2 but low quantities of IL-4 suggesting a combined Th1/Th2 cellular immune response. The vaccine administered either subcutaneously or orally protected mice against B. ovis infection. Such protection was similar to that provided by the reference living attenuated B. melitensis Rev. 1 vaccine. By contrast, only the subcutaneous vaccination with HS-PEC was as effective as Rev. 1 in conferring protection against B. abortus infection. The use of free HS or empty PEC microparticles did not produce any protective effect.

Abstract: The coming into effect on July 14 of the Zoonoses Order 1975, marks a new and important step in veterinary public health. The Order makes available powers to investigate and control Salmonella and Brucella infections in animals and Salmonella infections in birds which present a threat to human health. Any Salmonella and Brucella infection becomes reportable in certaain species when organisms have been isolated by laboratory test. Veterinary surgeons and livestock owners may be involved in this obligatory procedure. The statutory powers to investigate incidents and apply restrictions on the movement of animals, birds and their products are not limited to reportable species: they cover a much wider range of animals and birds. They will be held in reserve and used only when necessary because of a hazard to public health; the use of restrictions will be limited to cases involving a very serious threat to man. The information collected from reports and investigations will add considerably to the epidemiological knowledge of these infections and so allow improved methods of control to be developed. Some of these measures already in operation are described, together with others possible for the future.

Abstract: In 1970, voluntary State-based TB control programs in Australia were replaced by a coordinated national campaign to eliminate both brucellosis and tuberculosis from the cattle population. The campaign was funded and managed under tripartite agreement by State/Territory and Commonwealth governments and Industry. The tuberculosis component of the campaign relied on test and slaughter with surveillance for the disease in abattoirs and trace-back to property of origin an essential component. Because of the moderate sensitivity of the skin test ( approximately 70%), testing was repeated at prescribed intervals over a number of years. In the more hostile environment of northern Australia, novel strategies were developed to maximize musters and remove 'at risk' animals. Australia is fortunate it did not have a feral host for M. bovis (apart from buffalo, which were included in the campaign) to complicate eradication. A national granuloma submission program was implemented in 1992 to increase the intensity of abattoir monitoring. Selective or total depopulation was used in some herds to achieve the requirements of the national Standard Definitions and Rules of the Campaign and achieve the status of 'TB Free Area' in December 1997. Monitoring for tuberculosis has continued under the 5-year Tuberculosis Freedom Assurance Program and measures to further reduce the risk of new cases have been implemented.

Abstract: When considering an elk (Cervus elaphus) restoration program, wildlife managers must evaluate the positive and negative elements of translocation. We prepared this protocol to give an overview of health considerations associated with translocation of elk, with an emphasis on movement of free-ranging elk from western North America to the southeastern USA. We evaluated infectious agents and ectoparasites reported in elk from two perspectives. First, we made a qualitative estimate of the ability of the agent to be introduced and to become established. This was done using a selected set of epidemiologic factors. Second, if there was a good possibility that the organism could become established in the release area, the potential pathological consequences for elk and other wildlife, domestic animals, and humans were assessed via examination of the literature and consultation with other animal health specialists. The results of these evaluations were used to classify infectious agents and ectoparasites as low risk (n = 174), unknown risk (n = 10), and high risk (n = 9). We classified Anaplasma marginale, Anaplasma ovis, Mycobacterium paratuberculosis, Pasteurella multocida serotype 3, Elaphostrongylus cervi, Dicrocoelium dendriticum, Fascioloides magna, Echinococcus granulosus, Dermacentor albipictus, and Otobius megnini as unknown risks. High risk infectious agents and ectoparasites were the agent of chronic wasting disease, Brucella abortus, Mycobacterium bovis, Dermacentor andersoni, Ixodes pacificus, and Psoroptes sp. Parelaphostrongylus tennis, Elaeophora schneideri, and a Babesia sp. are parasites endemic in the southeastern USA that may present a "reverse risk" and adversely affect elk if released in some parts of the region. We developed a five-component protocol to reduce the risk of introduction of high risk infectious agents and ectoparasites that included: (1) evaluation of the health status of source populations, (2) quarantines, (3) physical examination and diagnostic testing, (4) restrictions on translocation of animals from certain geographic areas or populations, and (5) prophylactic treatment.

Abstract: Brucellosis is the zoonosis of world wide distribution and common cause of economic loss and ill health among animals and human populations. Patients with pyrexia of unknown origin (PUO) who were predisposed to brucellosis through rearing of animals and consumption of different animal products were tested for presence of Brucella abortus antibodies using Rose Bengal and serum agglutination antigens. Twenty six (5.2%) of the 500 patients had B. abortus antibody. The high titres of 320, 640 and 1280 obtained in the sera of patients in this study are suggestive of the endemicity of the disease in this environment. No significant difference in age and sex distribution of brucella antibody prevalence was observed. Similarly, spatial distribution of brucella antibody in different locations was not statistically significant. Although higher serological prevalence was noted in children and students than in other populations examined, the difference in prevalence between the various occupational groups was not significant. Animal handling activities including rearing are not important factors in the prevalence of brucellosis. However, among the rearers, the highest prevalence (20%) was observed among cattle handlers followed in decreasing order of prevalence by goat rearers (10%), mixed sheep and cattle rearers (9%), mixed sheep and goat rearers (8%), and 4% among each of sheep rearers and non rearers of animals. In addition, consumers of yoghurt and fresh goat milk had higher prevalence (20%) than consumers of other milk products. However, brucella antibody prevalence between consumers and non-consumers of animal products was not significantly different. The high economic loss and public health implications of brucellosis necessitates the need for effective surveillance as well as appropriate preventive and control measure among human and animal populations.

Abstract: Fluorescence polarization assay (FPA) is based on the rotational differences between a small soluble antigen molecule in solution (labelled with a fluorochrome) and the antigen molecule complexed with its antibody. A small molecule will rotate randomly at a rapid rate, resulting in rapid depolarization of light, while a larger complex molecule will rotate slower and depolarize light at a reduced rate. The rate change in depolarization can be measured. The FPA is a homogeneous assay which does not require removal of unreacted reagents and can, therefore, be performed very quickly and, given portable equipment, in the laboratory and in the field. The latter obviates the need for shipping samples and eliminates waiting for results, as well as reducing test costs.The FPA technology has been developed and validated for the serological diagnosis of brucellosis in cattle, swine, sheep, goats, bison, and cervids. Sufficient cross reactivity of the common epitopes of Brucella abortus, B. melitensis and B. suis O-polysaccharide (OPS) allowed for the use of a single antigen for all species of smooth Brucella and animals. The OPS prepared from B. abortus S1119.3 was conjugated with fluorscein isothiocyanate (FITC). The FPA was initially developed for testing serum; however, the technology has been extended to testing whole blood and milk from individual animals or bulk tank samples pooled from 2000 or fewer animals. The accuracy of the FPA equalled or exceeded those obtained using other serological tests such as the buffered antigen plate agglutination test (BPAT), the milk ring test (MRT), the complement fixation test (CFT), the indirect enzyme immunoassay (IELISA), and the competitive enzyme immunoassay (CELISA).

Abstract: A fluorescence polarization assay (FPA) for detection of antibody to Brucella abortus in individual milk samples was developed and validated. Samples from 190 cattle from which B. abortus was isolated; milk samples from cattle in herds infected with B. abortus (n = 1,086) and positive in the milk ring test (MRT), as well as milk samples from Canadian cattle (with no evidence of brucellosis, n = 2,974) were tested by the indirect enzyme immunoassay (IELISA) and the FPA. The sensitivity (based on samples from culture positive cattle) and specificity (based on Canadian milk samples) of the IELISA and the FPA were 100%. The relative sensitivity value obtained with milk from cattle of infected herds and the specificity values of the IELISA were 98.5 and 99.9%, respectively. The relative sensitivity and specificity of the FPA with the same samples were 82.2 and 99.4% using a cutoff value of 90 millipolarization units (mP). The low relative sensitivity value of the FPA was shown, by competitive enzyme immunoassay (CELISA), to be due to vaccinal antibody (assumed as vaccinal antibody against B. abortus Sl19 is excluded by the FPA and CELISA but not by the MRT and the IELISA), present in some of the milk samples. The FPA is a homogeneous assay which, unlike the MRT and the IELISA, may be used for testing in the field.

Abstract: OBJECTIVE: To determine the virulence of a Brucella abortus mutant, BA25, lacking a major 25 kd outer membrane protein (Omp25) in cattle. ANIMALS: 20 mixed-breed heifers in late gestation. PROCEDURE: 10 heifers were inoculated with 1 x 10(7) colony-forming units of the Omp25 mutant via the conjunctival sac, and an equal number were infected with the virulent parental strain B. abortus 2308. The delivery status of the dams was recorded, and colonization was assessed following necropsy. The ability of BA25 to replicate inside bovine phagocytes and chorionic trophoblasts was also evaluated in vitro because of the propensity of virulent brucellae to replicate inside these cells in vivo. RESULTS: The parental strain induced abortions in 5 of 10 inoculated cattle, whereas only 1 of 10 dams exposed to BA25 aborted. Brucella abortus strain 2308 colonized all of the cow-calf pairs and induced Brucella-specific antibodies in 100% of the dams. In contrast, BA25 was isolated by bacteriologic cultural technique from 30% of the calves and 50% of the inoculated dams (n = 10). Of the 10 heifers inoculated with BA25, 4 did not develop Brucella-specific antibodies nor were they colonized by the mutant strain. In bovine macrophages and chorionic trophoblasts, BA25 replicated in significantly lower numbers than the virulent parental strain (n = 3). CONCLUSIONS AND CLINICAL RELEVANCE: The 25 kd outer membrane protein may be an important virulence factor for B. abortus in cattle. The attenuation of the Omp25 mutant in cattle may involve the inability of BA25 to replicate efficiently in bovine phagocytes and chorionic trophoblasts.

Abstract: Serologic responses to the newly introduced rough Brucella abortus vaccine strain RB51 have been determined in a dot-blot format using gamma-irradiated RB51 cells as the antigen. Because gamma-irradiated cells are not easily prepared and the signal from cells was not always reliable, an alternative antigen was sought. Detergent extracts of B. abortus RB51 were prepared using zwittergent 3-14, Triton X-100, and sodium dodecyl sulfate (SDS) and examined in a dot-blot format. Zwittergent 3-14 extracts and gamma-irradiated RB51 cells gave the same titers. Unlike gamma-irradiated RB51 cells, zwittergent 3-14 extracts produced signals consistently, and the signals were easily interpreted. Triton X-100 extracts interfered with signal development, and SDS extracts resulted in a high background signal. Western blot analyses revealed several outer membrane proteins in the zwittergent 3-14 extract. The major antigens in the extract had apparent molecular weights of <20,000.

Abstract: Studies were carried out to detect the antibodies against zoonotic intracellular bacterial infections viz., brucellosis, listeriosis and tuberculosis in abattoir associated personnel employing dot-ELISA. Out of 165 serum samples tested 25.5, 40.0 and 10.9 per cent were detected as positive for brucellosis, listeriosis and tuberculosis, respectively. Immunodetection of these occupationally exposed persons for IgM as well as IgG antibodies revealed positivity for IgM, IgG and IgM + IgG in 8.5, 17.6 and 13.9 per cent for listeriosis and 5.5, 2.4 and 3.0 for tuberculosis respectively. Antibodies against brucellosis and listeriosis both were detected in 10.9 per cent persons, while 6.7 per cent persons were positive for both listeriosis and tuberculosis. No person was found positive with both brucellosis and tuberculosis. All the three infections were detected in 3.6 per cent persons.

Abstract: An enzyme immunoassay (EIA) was developed for detecting Brucella antibodies in milk of cows infected with Brucella abortus. The enzyme immunoassay using heat-killed cells of B. abortus strain 19 was of comparable sensitivity to the Brucella ring test in detecting antibodies in milk of a reference positive control cow experimentally infected with B. abortus strain 2308 and in milk of 16 naturally infected cows from which B. abortus was isolated. No detectable enzyme immunoassay reactions were present in milk of 11 noninfected controls. The enzyme immunoassay is a procedure which can be readily automated so that screening tests for brucellosis could be conducted at a reference laboratory where uniform conditions can be maintained.

Abstract: In six cases of Brucella melitensis infections the common etiologic factor was the ingestion of raw goat cheese. Presenting complaints were variable, but a constant feature was persistent fevers that frequently were present for months. Evidence of liver cell dysfunction was present in each case, and in two, biopsy disclosed diffuse hepatitis with focal necrosis. The diagnoses were initially based on high titers of Brucella agglutinins and later confirmed by positive blood cultures. A good clinical response was induced by antibiotic therapy ( tetracycline plus streptomycin sulfate), but in two cases a Jarish-Herxheimer reaction occurred during the first 24 hours of treatment.

Abstract: Three cases of brucellosis occurred, two acute cases and one chronic, all of which likely resulted from the ingestion of fresh cheese containing Brucella melitensis. A history of patients with illness suggestive of brucellosis should include details about food ingested recently in addition to the usual questions about travel and occupation. As the US domestic-animal disease comes under better control, and travel to Mexico, Spain, and Italy increases, it seems likely that human disease caused by B melitensis will become relatively more common.

Abstract: During the course of preliminary clinical trials of an improved membrane filter blood culture system, filter plugging produced by a gelling of the lysing solution was observed when the patients had high leukocyte counts. A solution of streptokinase-streptodornase (Varidase, Lederle Laboratories) dissolved or prevented the gel and permitted rapid filtration without plugging. With streptokinase-streptodornase incorporated in the filtration procedure, a comparison of several culture systems was carried out on 176 blood cultures. Brucella broth with and without sodium polyanethol sulfonate, a prereduced osmotically stabilized broth, pour plates, and an improved aerobic membrane filter system were compared. The membrane filter system yielded 29 of the total of 37 positive cultures, far surpassing all other systems. Eight of these cultures were detected first by the filter technique, and 13 were positive only in this system. Nineteen of the 37 positive cultures were from patients on antimicrobial agents. Fourteen of these were detected by the filter, twice the number detected by any of the other systems.

Abstract: The eradication of bovine tuberculosis and brucellosis of domestic stock in Germany through the combined efforts of veterinary medicine, the agricultural section and the state was an historic achievement. Since the two diseases are zoonoses, their successful control can also be seen as a valuable contribution to public health. Both these zoonoses are classic animal diseases presenting themselves as clinical entities complete with gross pathological lesions. In contrast, today we are confronted with pathogens causing zoonoses characterised by latent, i.e. clinically inapparent herd infections that do not result in visible tissue changes. Nevertheless, through contaminated foodstuffs, these pathogens contribute to food-borne infections leading to the outbreak of genuine zoonoses in humans. It has been estimated that there could be as many as two million cases of food-borne infections annually in Germany. Among them are salmonellosis, campylobacteriosis, yersiniosis, infections with verotoxin producing E. coli, listeriosis and toxoplasmosis. While the national animal disease legislation only foresees the control of notifiable diseases, the basis for zoonoses control is laid down in the EU Zoonosis-Directive, which is presently awaiting its transposition into national law and into practice. In order, for instance, to combat the most important Salmonella infections of humans, Integrated Quality Systems (IQS) have been formulated as a means of implementing the proven HACCP concept in animal production units and ensuring animal health from the point of view of consumer protection. The aim of all measures must be to free infected herds of pathogens, to investigate and eliminate all sources with a potential for further pathogen introductions, to maintain pathogen-free herds--with a reduced pathogen challenge in mid-term time periods--, as well as to develop diagnostics capable of identifying pathogen carriers before slaughter. For the disinfection of stock, it is important to have epidemiological data collecting systems and information systems that allow complete diagnostic tracing from herd to slaughterhouse and vice versa. All sides, including research and surveillance, as well as producers are called upon to actively share in protecting the health of consumers as far as it is threatened by latent infections in domestic stock.

Abstract: Employing the biparental exogenous plasmid isolation method, conjugative plasmids conferring mercury resistance were isolated from the microbial community of the rhizosphere of field grown alfalfa plants. Five different plasmids were identified, designated pSB101-pSB105. One of the plasmids, pSB102, displayed broad host range (bhr) properties for plasmid replication and transfer unrelated to the known incompatibility (Inc) groups of bhr plasmids IncP-1, IncW, IncN and IncA/C. Nucleotide sequence analysis of plasmid pSB102 revealed a size of 55 578 bp. The transfer region of pSB102 was predicted on the basis of sequence similarity to those of other plasmids and included a putative mating pair formation apparatus most closely related to the type IV secretion system encoded on the chromosome of the mammalian pathogen Brucella sp. The region encoding replication and maintenance functions comprised genes exhibiting different degrees of similarity to RepA, KorA, IncC and KorB of bhr plasmids pSa (IncW), pM3 (IncP-9), R751 (IncP-1beta) and RK2 (IncP-1alpha), respectively. The mercury resistance determinants were located on a transposable element of the Tn5053 family designated Tn5718. No putative functions could be assigned to a quarter of the coding capacity of pSB102 on the basis of comparisons with database entries. The genetic organization of the pSB102 transfer region revealed striking similarities to plasmid pXF51 of the plant pathogen Xylella fastidiosa.

Abstract: AIMS: To demonstrate the occurrence of cellulolytic bacteria in the termite Zootermopsis angusticollis. METHODS AND RESULTS: Applying aerobic cultivation conditions we isolated 119 cellulolytic strains from the gut of Z. angusticollis, which were assigned to 23 groups of aerobic, facultatively anaerobic or microaerophilic cellulolytic bacteria. 16S rDNA restriction fragment pattern and partial 16S rDNA sequence analysis, as well as numerical taxonomy, were used for the assignment of the isolates. The Gram-positive bacteria of the actinomycetes branch could be assigned to the order Actinomycetales including the genera Cellulomonas/Oerskovia, Microbacterium and Kocuria. The Gram-positive bacteria from the order Bacillales belonged to the genera Bacillus, Brevibacillus and Paenibacillus. Isolates related to the genera Afipia, Agrobacterium/Rhizobium, Brucella/Ochrobactrum, Pseudomonas and Sphingomonas/Zymomonas from the alpha-proteobacteria and Spirosoma-like from the "Flexibacteriaceae" represented the Gram-negative bacteria. CONCLUSIONS: A cell titre of up to 10(7) cellulolytic bacteria per ml, determined for some isolates, indicated that they may play a role in cellulose digestion in the termite gut in addition to the cellulolytic flagellates and termite's own cellulases. SIGNIFICANCE AND IMPACT OF THE STUDY: The impact of bacteria on cellulose degradation in the termite gut has always been a matter of debate. In the present survey we investigated the aerobic and facultatively anaerobic cellulolytic bacteria in the termite gut.

Abstract: Naturally acquired Brucella abortus infections were studied during consecutive pregnancies in eight sheep and in their lambs over a period of 40 months to evaluate epizootiologic aspects of natural infection in sheep. Brucella abortus was isolated from the ewes following 16 of 26 natural terminations of pregnancy: from 5 of 6 ewes in the first year, from six of eight ewes in the second year, from two of six ewes in the third year, and from three of six ewes in the fourth year. Vaginal swab samples and milk samples were the most consistent source of the brucella organisms. Brucella abortus was isolated from three ewes when standard tube test seroagglutination titers were less than 1:100. In contrast, results of supplemental tests (card, 2-mercaptoethanol, complement-fixation, and Rivanol) remained positive during the study. During the 40 months, B abortus was isolated from 4 of 4 aborted fetuses, 2 of 5 stillborn lambs, 10 of 37 living lambs, and as an indicator of continuing infection, from 6 of 12 lambs born during the fourth year. Although B abortus has a definite host preference for cattle, this study demonstrated that under appropriate management conditions, sheep may be naturally infected and may remain infected for more than 40 months. Epizootiologic evaluation of all factors, including husbandry practices and exposure potential, should be utilized in determining the need to test other species that may have been exposed to cattle infected with B abortus.

Abstract: Our analyses of lipopolysaccharide mutants of Sinorhizobium meliloti offer insights into how this bacterium establishes the chronic intracellular infection of plant cells that is necessary for its nitrogen-fixing symbiosis with alfalfa. Derivatives of S. meliloti strain Rm1021 carrying an lpsB mutation are capable of colonizing curled root hairs and forming infection threads in alfalfa in a manner similar to a wild-type strain. However, developmental abnormalities occur in the bacterium and the plant at the stage when the bacteria invade the plant nodule cells. Loss-of-function lpsB mutations, which eliminate a protein of the glycosyltransferase I family, cause striking changes in the carbohydrate core of the lipopolysaccharide, including the absence of uronic acids and a 40-fold relative increase in xylose. We also found that lpsB mutants were sensitive to the cationic peptides melittin, polymyxin B, and poly-l-lysine, in a manner that paralleled that of Brucella abortus lipopolysaccharide mutants. Sensitivity to components of the plant's innate immune system may be part of the reason that this mutant is unable to properly sustain a chronic infection within the cells of its host-plant alfalfa.

Abstract: Adverse effects of strain persistence and secretion in milk have been encountered with the Brucella melitensis vaccine strain Rev.1. Field isolates obtained from vaccinated animals and from a human resembled the vaccine strain Rev.1 by conventional bacteriological tests. The lack of a specific molecular marker that could specifically characterize the commercial vaccine strain prevented confirmation of the homology of the Rev.1-like field isolates to the vaccine strain. The composition of the omp2 locus from two gene copies with differences in their PstI restriction endonuclease sites was used to establish an epidemiologic fingerprint for the omp2 gene in the Rev.1 vaccine strain. Primers designed to amplify DNA sequences that overlap the PstI site revealed a single 282-bp DNA band common to all Brucella spp. Agarose gel electrophoresis of the PstI digests of the PCR products from strains 16M and the vaccine strain Rev.1 revealed a distinctive profile that included three bands: one band for the intact 282-bp fragment amplified from omp2a and two bands resulting from the digestion of the amplified omp2b gene fragment, 238- and 44-bp DNA fragments, respectively. Amplified fragments of 37 Rev.1-like isolates, including 2 human isolates, also exhibited this pattern. In contrast, DNA digests of all other Israeli field isolates, including atypical B. melitensis biotype 1 and representatives of the biotype 2 and 3 isolates, produced two bands of 238 and 44 bp, respectively, corresponding with the digestion of both omp2a and omp2b genes. This method facilitates identification of the Rev.1 vaccine strain in both animals and humans in Israel.

Abstract: The Dutch national Brucella abortus eradication programme for cattle started in 1959. Sporadic cases occurred yearly until 1995; the last infected herd was culled in 1996. In August 1999 the Netherlands was declared officially free of bovine brucellosis by the European Union. Before 1999, the programme to monitor the official Brucella-free status of bovine herds was primarily based on periodical testing of dairy herds with the milk ring test (MRT) and serological testing of all animals older than 1 year of age from non-dairy herds, using the micro-agglutination test (MAT) as screening test. In addition, serum samples of cattle that aborted were tested with the MAT. The high number of false positive reactions in both tests and the serum agglutination test (SAT) and complement fixation test (CFT) used for confirmation seemed to result in unnecessary blockade of herds, subsequent testing and slaughter of animals. For this reason, a validation study was performed in which three indirect enzyme-linked immunosorbent assays (ELISAs), the CFT and the SAT were compared using a panel of sera from brucellosis-free cattle, sera from experimentally infected cattle, and sera from cattle experimentally infected with bacteria which are known to induce cross-reactive antibodies (Pasteurella, Salmonella, Yersinia, and Escherichia). Moreover, four ELISAs and the MRT were compared using a panel of 1000 bulk milk samples from Brucella-free herds and 12 milk samples from Brucella abortus- infected cattle. It is concluded that the ELISA obtained from ID-Lelystad is the most suitable test to monitor the brucelosis free status of herds because it gives rise to fewer false-positive reactions than the SAT.

Abstract: AIMS: To select an anti-fungal agent to replace cycloheximide in the media used for isolation of Brucella. METHODS AND RESULTS: One potential agent, natamycin, was evaluated using 28 Brucella isolates, 18 yeasts and 14 fungi. The material for the evaluation included 37 bovine milk samples, six bovine vaginal swabs and 45 milk samples artificially infected with Brucella. The recovery of Brucella only from the artificially-inoculated milk samples increased with the use of the medium containing natamycin instead of cycloheximide, at the same time significantly inhibiting the growth of yeasts, fungi and other bacteria. The inclusion of either anti-fungal agent allowed growth of the 28 Brucella isolates and totally prevented the growth of all 18 yeasts and 13 of the 14 fungi. CONCLUSIONS: Based on the results it was concluded that natamycin would be a suitable alternative to cycloheximide. SIGNIFICANCE AND IMPACT OF THE STUDY: Cycloheximide has become unavailable worldwide and is currently an anti-fungal constituent of the medium often used for isolation of Brucella organisms. The use of natamycin as a replacement in the formulation did not inhibit growth of Brucella and was effective at eliminating most contaminants.

Abstract: STUDY DESIGN: Case report. OBJECTIVE: To increase awareness of spinal brucellosis and discuss demographics, diagnosis, and treatment. SUMMARY OF BACKGROUND DATA: Brucellosis is a rare cause of spinal infections in the United States, although there have been regional increases in its prevalence. METHODS: Retrospective review of a patient with spinal brucellosis. She underwent a protracted course of treatment, with a long delay in diagnosis. History ultimately revealed regular consumption of unpasteurized goat cheese. Appropriate testing subsequently led to the diagnosis. RESULTS: After multiple surgeries and medications, the condition responded well to definitive antibiotic therapy. CONCLUSION: Brucellosis is rare in the United States and thus often overlooked in the differential diagnosis of back pain. The changing risk pattern for this disease requires a high index of suspicion, which can result in early diagnosis and predictably favorable results to treatment.

Abstract: OBJECTIVE: This study was performed to determine the ability of the VITAL system to detect and allow recovery of microorganisms that are difficult to grow, such as Brucella spp., yeasts, or anaerobes, as well as to determine the need for blind subcultures after the incubation period. METHODS: A prospective evaluation of the system was performed, and 8247 blood culture bottles were processed. The standard was blind subculture from all the bottles after 5 days of incubation. RESULTS: There were 3.2% false-positive and 0.6% false-negative results (72% of clinical importance). The system sensitivity for yeasts was 41%. The mean time for detection of Neisseria meningitidis was 31.9 +/- 2.8 h, for Brucella spp. 119.7 +/- 2 h, and for yeast 51.5 +/- 27.8 h. CONCLUSIONS: The VITAL system poses has serious difficulties in the detection of N. meningitidis, Brucella spp., yeast and methicillin- and aminoglycoside-resistant Staphylococcus aureus (MARSA). The low system sensitivity for yeast detection makes the blind subculture necessary after the incubation period.

Abstract: Plasmid pIPO2 is a cryptic, conjugative, broad-host-range plasmid isolated from the wheat rhizosphere. It efficiently self-transfers between alpha, beta and gamma Proteobacteria and has a mobilizing/retromobilizing capacity for IncQ plasmids. The complete nucleotide sequence of pIPO2 is presented on the basis of its mini-Tn5::luxABtet-tagged derivative, pIPO2T. The pIPO2 sequence is 39815 bp long and contains at least 43 complete ORFs. Apart from a suite of ORFs with unknown function, all of the genes carried on pIPO2 are predicted to be involved in plasmid replication, maintenance and conjugative transfer. The overall organization of these genes is different from previously described plasmids, but is similar to the genetic organization seen in pSB102, a conjugative plasmid recently isolated from the bacterial community of the alfalfa rhizosphere. The putative conjugative transfer region of pIPO2 covers 23 kb and contains the genes required for DNA processing (Dtr) and mating pair formation (Mpf). The organization of these transfer genes in pIPO2 is highly similar to the genetic organization seen in the environmental plasmid pSB102 and in pXF51 from the plant pathogen Xylella fastidiosa. Plasmids pSB102 and pXF51 have recently been proposed to form a new family of environmental broad-host-range plasmids. Here it is suggested that pIPO2 is a new member of this family. The proposed Mpf system of pIPO2 shares high amino acid sequence similarity with equivalent VirB proteins from the type IV secretion system of Brucella spp. Sequence information was used to design primers specific for the detection of pIPO2. Environmental DNA from a range of diverse habitats was screened by PCR with these primers. Consistently positive signals for the presence of pIPO2 were obtained from a range of soil-related habitats, including the rhizospheres of young wheat plants, of field-grown oats and of grass (all gramineous plants), as well as from the rhizosphere of tomato plants. These data add to the growing evidence that plasmids carry advantageous genes with as yet undefined functions in plant-associated communities.

Abstract: Microparticles, containing an antigenic complex from Brucella ovis (HS), were evaluated for vaccine purposes against brucellosis. They were prepared by the double emulsion solvent evaporation method using two different polyesters, poly-lactide-co-glycolide acid (75:25; RG 756) and poly-epsilon-caprolactone. The encapsulation efficiency and release of HS from the microparticles, their capacity to be phagocytosed and also their toxicity on murine monocytes J774.2 were investigated. Both polymers lead to smooth and spherical sub-5 microm particles, with approximately 30% of the antigen initial dose encapsulated. SDS-PAGE and immunoblot of extracted antigens confirmed that the apparent molecular weight and antigenicity remained unaltered after the encapsulation procedure. However, the in vitro release of the antigens differed among them. The release profile for PLGA microparticles was continuous, whereas PEC ones released the antigens in a triphasic release pattern. Phagocytosis was clearly influenced by the hydrophobicity of the polymer, increasing in the case of PEC microparticles. Toxicity assay showed that both types of microparticles induced similar levels of mitochondrial damage. In conclusion, HS-PEC microparticles could be used as an effective vaccine against brucellosis, as the antigen is released in boosters and they are greatly phagocytosed by macrophages.

Abstract: One hundred and three milk samples were collected from 52 cows, 21 ewes, 18 goats and 12 camels. The animals tested positive to at least one of the following: (1) standard tube agglutination test (SAT); (2) Rose Bengal plate test (RBPT); (3) milk ring test (MRT). All milk samples were examined by culture and single-step polymerase chain reaction (PCR) techniques for detection of Brucella species. The PCR assay amplified Brucella-DNA from 29 bovine milk samples, 10 from sheep, 13 from goats and one from a camel. The direct culture method detected Brucella organisms from 24 samples of cows' milk, 12 from sheep, 10 from goats and failed to detect any Brucella organisms from camels' milk. PCR detected up to 100 colony forming units (CFU) of B. abortus per millilitre of milk in 100% of diluted milk samples, and 1000 CFU of B. melitensis from 70% of milk samples. Although the overall sensitivity of the PCR was higher than the culture method, it should be possible to increase the sensitivity to detect lower numbers of Brucella organisms in field samples. The speed and sensitivity of the PCR assay suggest that this technique could be useful for detection of Brucella organisms in bovine milk, as well as in sheep, goat, and camels milk.

Abstract: An automated indirect enzyme-linked immunosorbent assay (I-ELISA) for the serological diagnosis of bovine brucellosis was developed and validated in-house. A total of 4,803 cattle sera from South Africa (n = 3,643), Canada (n = 652), Germany (n = 240), France (n = 73) and the USA (n = 195) was used. The South African panel of sera represented 834 sera known to be positive by the Rose Bengal test (RBT), serum agglutination test (SAT) and complement fixation test (CFT), 2709 sera that were negative by CFT, and 100 sera from animals vaccinated with a standard dose of Brucella abortus strain 19. Overseas sera were obtained from reference non-vaccinated brucella-free cattle (n = 834), naturally infected (n = 72), experimentally infected (n = 71), and vaccinated animals (n = 83). Also 100 sera collected from cattle in Canada and known to be positive by competitive ELISA (C-ELISA) were used. The intermediate ranges ("borderline" range for the interpretation of test results) were derived from two-graph receiver operating characteristics analysis. The lowest values of the misclassification cost-term analysis obtained from testing overseas panels, covered lower I-ELISA cut-off PP values (0.02-3.0) than those from local panels (1.5-5.0). The relatively low cut-off PP values selected for I-ELISA were due to the fact that the positive control used represents a very strong standard compared to other reference positive sera. The greater overlap found between negative and positive cattle sera from South Africa than that between reference overseas panels was probably due to the different criteria used in classifying these panels as negative (sera from true non-diseased/non-infected animals) or positive (sera from true diseased/infected animals). The diagnostic sensitivity of the I-ELISA (at the optimum cut-off value) was 100% and of the CFT 83.3%. The diagnostic specificity of I-ELISA was 99.8% and of the CFT 100%. Estimate of Youden's index was higher for the I-ELISA (0.998) than that for the CFT (0.833). Analysis of distribution of PP values in sera from vaccinated and naturally infected cattle shows that in vaccinated animals all readings were below 31 PP where in infected ones these values represented 43%. Therefore, it appears that I-ELISA could be of use in identifying some naturally infected animals (with values > 31 PP), but more sera from reference vaccinated and infected animals need to be tested to further substantiate this statistically. Of 834 sera positive by RBT, SAT and CFT, 825 (98.9%) were positive in the I-ELISA. Compared to C-ELISA the relative diagnostic sensitivity of the I-ELISA was 94% and of the CFT 88% when testing 100 Canadian cattle sera. Of 258 South African cattle sera, of which 183 (70.9 %) were positive by the I-ELISA and 148 (57.4 %) by the CFT, 197 (76.4%) were positive by C-ELISA when re-tested in Canada. One has to stress, however, that Canadian C-ELISA has not been optimised locally. Thus, the C-ELISA was probably not used at the best diagnostic threshold for testing South African cattle sera. This study shows that the I-ELISA performed on an automated ELISA workstation provides a rapid, simple, highly sensitive and specific diagnostic system for large-scale detection of antibodies against B. abortus. Based on the diagnostic accuracy of this assay reported here, the authors suggest that it could replace not only the currently used confirmatory CFT test, but also the two in-use screening tests, namely the RBT and SAT.

Abstract: Infection with Brucella spp. continues to pose a human health risk in California despite great strides in eradicating the disease from domestic animals. Clustering of human cases in time and space has important public health implications for understanding risk factors and sources of infection. Temporal-spatial clustering of human brucellosis in California for the 20-year period 1973-1992 was evaluated by the Ederer-Myers-Mantel, Moran's I, and population-adjusted Moran's I procedures. Cases were clustered in concentrated agricultural regions in the first 5-year interval (1973-1977). Time-space clustering of human brucellosis cases in California late in the 20-year study period may reflect the distribution of Hispanic populations. Public health programs in California should focus on educating Hispanic populations about the risk of consuming dairy products, such as soft cheeses, made from unpasteurized milk.

Abstract: Campylobacter jejuni is a thermophilic and microaerophilic enteric pathogen associated with poultry. Biofilms may be a source of C. jejuni in poultry house water systems since they can protect constituent microorganisms from environmental stress. In this study, the viability of C. jejuni in biofilms of gram-positive chicken house isolates (P1, Y1, and W1) and a Pseudomonas sp. was determined using a cultural method (modified brucella agar) and direct viable count (DVC). Two-day biofilms grown on polyvinyl chloride (PVC) coupons in R2A broth at 12 and 23 degrees C were incubated with C. jejuni for a 6-h attachment period. Media were then refreshed every 24 h for 7 days to allow biofilm growth. Two-day biofilms of P1, Y1, and Pseudomonas spp. enhanced attachment (P < 0.01) of C. jejuni (4.74, 4.62, and 4.78 log cells/cm2, respectively) compared to W1 and controls without preexisting biofilm (4.31 and 4.22 log cells/cm2, respectively). On day 7, isolates P1 and Y1 and Pseudomonas biofilms covered 5.4, 7.0, and 21.5% of the surface, respectively, compared to 4.9% by W1. Viable C. jejuni on the surface decreased (P < 0.05) with time, with the greatest reduction occurring on surfaces without a preexisting biofilm. The number of viable C. jejuni determined by DVC was greater than that determined by the cultural method, indicating that C. jejuni may form a viable but nonculturable state within the biofilm. Both DVC and the cultural method indicate that biofilms enhance (P < 0.01) the survival of C. jejuni during incubation at 12 and 23 degrees C over a 7-day period.

Abstract: A seroepidemiological study carried out in a high-risk village in Crete in 1985-1987 and 1998 showed that although the awareness of the people concerning zoonoses had increased during this period, the situation did not improve: there was a significant increase of the spread of seroprevalence in time and space of Coxiella burnetii, Rickettsia typhi, Brucella sp., and Entamoeba histolytica. Toxoplasma gondii, Rickettsia conorii, Borrelia burgdorferi, Echinococcus granulosus, Leishmania sp., and Fasciola hepatica stayed at the same levels. This first study of Bartonella henselae in Crete showed that 15.9% of the children tested were seropositive. The results indicate that reservoirs and vectors of the pathogens studied are widespread in the environment, and the way of life of the people favors contact with them. Seven of 30 milk samples were positive for Brucella sp. by seminested polymerase chain reaction.

Abstract: In this study, a total of 105 samples including 35 raw milk, 35 cows' milk cheese, and 35 ewes' milk cheese samples were obtained from Kirikkale city and investigated for the presence and contamination level of Brucella species. For this reason, milk and cheese samples were collected in aseptic conditions and brought to the laboratory under cold conditions. The method suggested from FARREL was used for the isolation and identification of Brucella spp. In addition, Most Probable Number Technique was used to determine the contamination level in Brucella spp. positive samples. According to analysis findings, B. melitensis was isolated from 5 (14.2%) of 35 ewes' milk cheese samples at the level of 3.6 x 10(1)-9.3 x 10(3) MPN/g. Brucella spp. were not detected in any of raw milk and cows' milk cheese samples. In conclusion this study reinforced the previous reports that ewes' milk cheese is an important source of Brucella spp. and they have been risk for public health.

Abstract: The authors present the results of study of the antibody neutralization test in brucellosis with the use of erythrocytes sensitized with lipopolysaccharide. This test was shown to be highly specific and sensitive in detection of brucellae and brucellosis antigen in pathological material, food and objects of external environment.

Abstract: The antigenic extract Hot Saline from Brucella ovis was microencapsulated by the spray-drying technique with different polyesters (poly-lactide-co-glycolide RG502H [PLGA], and blends with poly- epsilon -caprolactone [PEC]) in order to obtain microparticles smaller than 5 microm. Microparticles were tested for encapsulation efficiency, release studies, acidification of the in vitro release medium, and in vitro J744-macrophage experiments (phagocytosis and toxicity of the preparations) to determine the optimal formulation for vaccination purposes. Formulation containing no PCL showed the highest encapsulation efficiency, although the differences were not significant. The in vitro release kinetics were characterized by a high burst effect after 1 h of incubation, followed by a slow and continuous release. For the formulation based on PLGA, the pH of the medium during release dropped from 7.4 to 3.5 while the presence of PEC attenuated the pH drop. All formulations showed light toxicity by the MTT assay, but differences were observed in terms of phagocytosis, as particles prepared with PEC showed the higher uptake by J744-macrophages and cell respiratory burst, determined by oxygen peroxide release. All these characteristics suggest that the microparticulated antigenic formulation containing the higher ratio of PEC is susceptible to be used in animal vaccination studies.

Abstract: Autoinducer molecules are utilized by gram-negative and gram-positive bacteria to regulate density-dependent gene expression by a mechanism known as quorum sensing. PCR and DNA sequencing results showed that Campylobacter jejuni and Campylobacter coli possessed luxS, which is responsible for autoinducer-2 (AI-2) production. Using a Vibrio harveyi luminescence assay, the production of AI-2 was observed in milk, chicken broth, and brucella broth by C. coli, C. jejuni, Salmonella enterica serovar Typhimurium, and Escherichia coli O157:H7 under different conditions.

Abstract: An epidemiological survey was carried out in an area where there is an abattoir/canning factory in which a recrudescence of cases of professional brucellosis was observed: whilst there had been 29 cases of brucellosis in 8 years, 10 cases were confirmed in 1970. Our survey had underlined a brucellosis endemy in the population of that area: 32.50% of the patients had agglutinating or complement-fixing antibodies (698 people were examined). Results obtained have brought us to discuss the professional origin of brucellosis.

Abstract: A tremendous volcanic eruption destroyed all the life around Mount Vesuvius during the night between 24 and 25 August, 79 AD. Two famous towns, Pompeii and Herculaneum, were completely buried under volcanic products. At Herculaneum, about 25m of volcanic mud killed about 250 people who had fled to the beaches in an attempt to escape (Bisel, S. C.,Rivista di Studi Pompeiani, 1, 123-124, 1987). An anthropological examination of the skeletons of these "fugitives" reveals the bone lesions typical of brucellosis in 17.4% of adults (Capasso, L., International Journal of Osteoarchaelogy, 9, 277-288, 1999). This very high incidence of brucellosis was theoretically linked to the consumption of ovine milk and its derivates, which is also indicated by both literary and figurative sources. A single carbonized cheese was found in Herculaneum; its analysis clearly reveals the excellent state of preservation of the milk curds. For the first time, we demonstrate the presence of a variety of bacteria, possibly Lactobacillus, that also includes cocco-like forms that seem to be morphologically and dimensionally consistent with Brucella. The long interval spent by the organic remains under the volcanic mud and high temperatures they suffered preclude the possibility of identifying the bacteria through molecular methods.

Abstract: Fluorescence polarization assay (FPA) is a homogeneous technique which was applied to the serological diagnosis of bovine brucellosis. Because of its simplicity and because it may be performed very rapidly, it was an ideal test to adapt to field use. The FPA was used to test cattle on six dairy farms in Baja California, Mexico. Anticoagulated blood, serum, and milk were collected from each animal. The anticoagulated blood was tested immediately on the farm while serum and milk were tested subsequently in the laboratory. Cattle on one farm (n = 140) were thought not to be infected with Brucella abortus and the other farms were thought to have high prevalence of the infection. The whole blood FPA (FPA(bld)) did not detect antibody in any of the cattle on the first premise. This finding was confirmed using a number of other serological tests, including the buffered antigen plate agglutination test, the complement fixation test, the indirect and competitive enzyme immunoassays, and the FPA using serum and milk. Cattle on the other premises (n = 1122) were tested in a similar fashion. The sensitivity of the FPA(bld), relative to the serum FPA (considered the definitive test), was 99.1% and the relative specificity of the FPA(bld) was 99.6%. These results compared favourably with those obtained using the other serological tests.

Abstract: The antimicrobial activity of substances used as antibacterial agents (solutions of 10% calcium hydroxide, camphorated paramonochlorophenol - PMCC, 2% chlorhexidine digluconate and 10% castor oil plant detergent) on anaerobic bacteria (Fusobacterium nucleatum ATCC 25586, Prevotella nigrescens ATCC 33563, Clostridium perfringens ATCC 13124 and Bacteroidesfragilis ATCC 25285), using a broth dilution technique, was evaluated in vitro. For determination of minimum inhibitory and minimum bactericide concentrations (MIC and MBC), two culture broths, Reinforced Clostridial Medium (RCM) and supplemented Brucella, standardized inoculum and serially diluted solutions were used. All antibacterial agents presented antimicrobial activity that varied for different bacteria. There were no differences in the performance of the two broths. Chlorhexidine digluconate was the most effective, with the lowest MICs, followed by castor oil detergent, PMCC and calcium hydroxide. C. perfringens and B. fragilis were the most resistant bacteria to all agents.

Abstract: The BacA protein is essential for the long-term survival of Sinorhizobium meliloti and Brucella abortus within acidic compartments in plant and animal cells, respectively. Since both the S. meliloti and B. abortus bacA mutants have an increased resistance to bleomycin, it was hypothesized that BacA was a transporter of bleomycin and bleomycin-like compounds into the bacterial cell. However, our finding that the S. meliloti bacA mutant also has an increased sensitivity to detergents, a hydrophobic dye, ethanol, and acid pH supported a model in which BacA function affects the bacterial cell envelope. In addition, an S. meliloti lpsB mutant that is defective at a stage in infection of the host similar to that found for a bacA mutant is also sensitive to the same agents, and the carbohydrate content of its lipopolysaccharide (LPS) is altered. However, analysis of crude preparations of the bacA mutant LPS suggested that, unlike that for LpsB, BacA function did not affect the carbohydrate composition of the LPS. Rather, we found that at least one function of BacA is to affect the distribution of LPS fatty acids, including a very-long-chain fatty acid thought to be unique to the alpha-proteobacteria, including B. abortus.

Abstract: The rapid automated bacterial impedance technique (RABIT) was examined as a method for the detection of two wild-type isolates of Campylobacter coli in broth media. Both isolates failed to produce a change in impedance that was sufficient for detection in any combination of six nonselective basal broth media, including Mueller-Hinton broth, nutrient broth no. 2, brain heart infusion broth supplemented with yeast extract (0.5% [wt/vol]), brucella broth, Campy broth supplemented with yeast extract (0.5% [wt/voll), and Whitley impedance broth, at 37 and 42 degrees C. Although the strains did proliferate in the media, changes in conductivity were very small (ranging from 0 to 1,000 microS) and were not significantly greater than the drift in conductance observed in the control broth medium. Additional work is therefore required to define a nonionic growth substrate that will produce charged ions upon metabolism that are detectable by RABIT.

Abstract: The role of a few microorganisms, like Brucella and Mycobacterium and certain parasites of food animals, in causing human disease has been recognized for a hundred years. By the 1990s, other microorganisms derived from food animals were recognized as contributing to human illness. Handling and/or consumption of wild game may result in human exposure to novel microorganisms; these unrecognized or unknown agents or diseases in wild species may cross into humans and cause "new" diseases with which we are not familiar.

Nishi JS, Stephen C, Elkin BT. Implications of agricultural and wildlife policy on management and eradication of bovine tuberculosis and brucellosis in free-ranging wood bison of northern Canada. Annals of the New York Academy of Sciences. 2002 Oct; 969; 236-44. [PubMed: 12381598].

Abstract: Although disease is often an important factor in the population dynamics of wild ungulates, it is largely the threat-both real and perceived-that sylvatic disease reservoirs pose to the health status of commercial livestock or game farm industry that has led governments to establish policy and legislation for disease management, trade, and movement. With respect to bovine tuberculosis and brucellosis in wildlife, policies are largely borrowed from the existing regulatory framework for domestic livestock. In this paper, we review how general policy goals for managing these reportable diseases in domestic livestock have also affected conservation and management of bison in Canada. We argue that there is a need to better integrate conservation biology with agricultural livestock policy to develop management options and better address the unique conservation challenges that diseased free-ranging bison populations present.

Abstract: The process of Bovine Brucellosis Eradication that began in 1996 in the 10th Region de Los Lagos of Chile will be reviewed. The region comprises the most important dairy area of the country and it has the largest concentration of brucellosis infected herds. Based on the information gathered by an epidemiological surveillance system, the results of the eradication process for the years 1996 till 2001 are presented as rates of Milk Ring Test (MRT) positive dairies, rates of brucellosis reactors (bovines) in livestock markets and slaughterhouses, and the annual incidence and prevalence of brucellosis infected herds. During the period the rates of positive dairies, bovine reactors in livestock markets and slaughterhouses, and the annual incidence and prevalence of infected herds have experienced a decrease, while the rate of bovine reactors in slaughterhouses has remained stable. Data on the preventive measures taken, such as vaccination of female bovines and Certification of Brucellosis Free Herds, are also shown. The surveillance system has allowed the detection of infected herds, while the measures of prevention and cleaning of infected herds have allowed a reduction in the incidence and prevalence of the infection by Brucella abortus.

Abstract: A short review of the brucellosis situation, its control and eradication programs are presented. Data from over 1.2 million samples collected from more than 50,718 groups of cattle over a period of over 20 years (1979-2000) illustrates that over the last few years the number of individual reactors remain constant at around 3-4%. The percentage of reactive groups of animals decreased over these years, reflecting a better disease management and possibly an improved general education, handling of information on the immune (vaccination) status of animals and testing practices. Reported zoonotic cases are presented, as well as control and eradication programs, including utilization of vaccines.

Abstract: Brucellosis has been recognized in Argentina since the 19th century. Several studies demonstrated the presence of the disease in most of the domestic species. Actually, the estimate of prevalence is that between 10 and 13% of the farm animals are infected with bovine brucellosis with an individual rate of 4-5%. The annual economical losses have been estimated at 60,000,000 US dollars. The control of bovine brucellosis began in 1932 and successive resolutions have been issued since then. The current resolution indicates that B. abortus S19 is mandatory in female calves between 3 and 8 months of age. The vaccine strain B. abortus RB51 was provisionally approved but only for cattle older than 10 months of age. The brucellosis control program consists principally of test and slaughter. This methodology has been successful mainly in the dairy farms that have the incentive due to increased pricing because of obtaining a low prevalence of the disease. Brucellosis has been found in porcine, caprine, ovine and canine species. All Brucella species have been found in the country. Human brucellosis is an important disease and a national coordinated diagnostic net has been formed to better control the disease in man.

Abstract: Brucellosis is an important disease among livestock and people in sub-Saharan Africa. In general, the incidence is the highest in pastoral production systems and decreases as herd size and size of landholding decreases. The prevalence of risk factors for infections are best understood for bovine brucellosis and to a lesser extent for ovine and caprine brucellosis. The occurrence and epidemiology of brucellosis in pigs is poorly understood. This species bias is also reflected in control activities. As with other public-sector animal health services, the surveillance and control of brucellosis in sub-Saharan Africa is rarely implemented outside southern Africa. Brucellosis is even more ignored in humans and most cases go undiagnosed and untreated, leading to considerable suffering for those affected. Decision-making to determine the importance of brucellosis control relative to other public concerns and what brucellosis control strategies should be applied is urgently required. A strategy for how brucellosis decision-making might be considered and applied in future is outlined.

Abstract: This article reports epidemiological investigations on the occurrence of brucellosis in Romania. Like in other former communist countries, data concerning epidemiology of brucellosis and published articles are very few. The epidemiology and control of brucellosis in Romania was analyzed using data made available by the Office International des Epizooties and Veterinary Service of Romania. Romania, like many other developed countries, has eradicated Brucella abortus from cattle since 1969. Brucellosis caused by Brucella melitensis has never been reported. The incidence of brucellosis in swine and sheep is very rare but still there are a few outbreaks in some regions. In 2000, the number of cases was 47 in swine and 270 cases in sheep. Vaccination against brucellosis is prohibited in Romania.

Abstract: Infection by Brucella abortus seems to be a major cause of abortions among cattle and buffaloes in Sri Lanka. The incidence of this disease is more prominent among the animals in the Dry zone of the country raised under extensive management systems. The present low incidence of this disease and the small size of the country may facilitate launching of an effective disease control scheme. The milk ring test (MRT) has proven to be usable in testing milk for the infection at farm level. An ELISA technique could be employed to test the seroprevalence of infection among MRT-positive animals. A program to purchase the diseased animals by the state for slaughter, and a countrywide vaccination program with B. abortus strain RB51 would enable the country's livestock industry to eventually eradicate this disease.

Abstract: Serological diagnosis of brucellosis began more than 100 years ago with a simple agglutination test. It was realized that this type of test was susceptible to false positive reactions resulting from, for instance, exposure to cross reacting microorganisms. It was also realized that this test format was inexpensive, simple and could be rapid, although results were subjectively scored. Therefore, a number of modifications were developed along with other types of tests. This served two purposes: one was to establish a rapid screening test with high sensitivity and perhaps less specificity and a confirmatory test, usually more complicated but also more specific, to be used on sera that reacted positively in screening tests. This led to another problem: if a panel of tests were performed and they did not all agree, which interpretation was correct? This problem was further compounded by the extensive use of a vaccine which gave rise to an antibody response similar to that resulting from field infection. This led to the development of an assay that could distinguish vaccinal antibody, starting with precipitin tests. These tests did not perform well, giving rise to the development of primary binding assays. These assays, including the competitive enzyme immunoassay and the fluorescence polarization assay are at the apex of current development, providing high sensitivity and specificity as well as speed and mobility in the case of the fluorescence polarization assay.

Abstract: A simple, rapid, inexpensive fluorescence polarization assay for the detection of antibodies to Brucella abortus in bulk tank milk samples at the farm level or at dairies with a sensitivity and specificity of 100 and 95.9%, respectively, is described. The assay detects antibodies to B. abortus in 15 min by testing undiluted whey produced by chemical and physical manipulation of milk from bulk tanks. This sampling is noninvasive and therefore costs less and is less stressful than blood-based tests. The assay is specific and can detect antibodies at levels below that of the indirect enzyme immunoassay for milk and the fluorescence polarization assay for individual milk samples. Use of this test would make programs for surveillance of dairy animals and eradication of B. abortus more cost-effective.

Abstract: This study was conducted to evaluate the ability of the BacT/Alert automated blood culture system to detect Brucella spp. in comparison with traditional Brucella broth culture. Overall, 100 (50 bone marrow and 50 blood samples) paired cultures were obtained, and 59 were positive by at least one method. The Brucella broth culture method detected all 59 positive cultures (100%), and the BacT/Alert system detected 30 (50.8%) (P < 0.05). The mean detection times for B. melitensis were 4.5 days in the BacT/Alert system and 5 days in Brucella broth culture (P > 0.05). There is no significant difference between the two methods with respect to growth time of the microorganism, but Brucella broth culture is more sensitive than the BacT/Alert system.

Abstract: The diagnosis of brucellosis is generally made when a standard tube agglutination titer of 1/160 or more for anti-Brucella antibodies in the presence of compatible clinical signs and symptoms. However isolation of the organism from blood or bone marrow is the proof of the disease. In this study we aimed to describe the rate and duration of isolation of Brucella spp. from blood and bone marrow by use of automated blood culture system (BACTEC 9240). Between 1997 to 2001, 23 adults were diagnosed as brucellosis. Blood culture was obtained in all and simultaneous bone marrow culture in 18 and both specimens were cultured by BACTEC 9240. Brucella was isolated from blood and bone marrow cultures in 19 (82.6%) and 13 (81.2%) respectively. All positive blood cultures yielded within 7 days and bone marrow cultures in 4 days. We concluded that automated BACTEC culture systems can isolate Brucella spp. in a fast and efficient way.

Abstract: A description of some of the clinical features of low back pain in the horse has been given and a number of methods for assisting diagnosis considered. As well as a complete clinical examination both at rest and during exercise, a useful diagnostic aid in some chronic cases was the injection of local anaesthetic into the interspinous spaces. A laboratory examination, including haematological and biochemical profiles, was undertaken in all cases. The serum enzymes GOT and CPK were particularly valuable as an aid to diagnosis in atypical tying-up. A technique for radiography of the vertebral column of the mid back in the standing position and of the pelvic and sacroiliac regions of the anaesthetised horse was described using a Siemens Triplex Optimatic 1023 machine. Some of the radiological features of the vertebral column were considered and a breakdown of the diagnosis of 110 referred back cases. The most important conditions included muscle strain, crowding and overriding of the dorsal spinous processes in the mid back, spondylosis, undue curvature of the spine and vertebral fractures.

Abstract: Examination of hospital and public health records revealed 19 cases of brucellosis diagnosed in Philadelphia between 1968 and 1972. A serologic survey at Philadelphia's largest hog-processing plant, however, indicated infection in 39% of workers. If extrapolated industry-wide, the total would be several hundred in Philadelphia. The infection is usually unrecognized or asymptomatic, since men were active in physically demanding jobs with agglutinin titers in excess of 1:5000. Overt illness, usually first diagnosed after weeks of incapacity, responded readily to tetracycline therapy. Although clinical manifestations are nonspecific, attention to occupational history should quickly lead to the diagnosis. It is emphasized that any hog-processing plant, wherever located, is potentially a reservoir of brucellosis. Prolonged morbidity and loss of production time might be avoided if physicians were more alert to this infection. Agglutinins in possibly significant titers were also found in a small fraction of persons without identifiable exposure.

Abstract: An indirect enzyme-linked immunosorbent assay (ELISA) was developed to identify elk (Cervus elaphus nelsoni) with Brucella abortus strain RB51 (RB51)-specific antibodies using a mouse monoclonal antibody specific for bovine IgG1. This test was relatively easy to perform, accurate, and easily reproducible; therefore it could be standardized for use between laboratories. In addition, we attempted to compensate for inherent variabilities encountered when comparing ELISA readings from multiple samples taken from many animals over time. Optical density (OD) readings for each sample were converted into a percent positivity value for analysis. A negative cutoff value was determined above which a sample was considered to have a significantly elevated anti-RB51 antibody level. Pre- and postvaccination sera from 64 6-8 mo old elk, divided into four groups (females subcutaneously inoculated with saline (control animals), females ballistically inoculated with RB51, females subcutaneously inoculated with RB51, and males subcutaneously inoculated with RB51) were used. All serum samples were collected between 27 April and 15 November 1995. Values for all saline controls were appropriately below the negative cutoff value. All subcutaneously and ballistically inoculated elk were serologically positive to RB51 for at least two sampling periods during the study. The difference in percent positivity values for the ballistically compared to the subcutaneously inoculated groups was not statistically significant at 8, 10, 14, or 18 wk postvaccination. This suggests that processing RB51 into lactose based pellets and ballistically inoculating elk with these pellets does not alter the detectable elk antibody response. Also, inoculated and control animals can be accurately identified with ELISA at 4-8 weeks postvaccination.

Abstract: This prospective study was carried out in the department of infectious diseases of Gaziantep University between January 1997 and December 1999 to evaluate the epidemiological, clinical and laboratory features of brucellosis in south-eastern Turkey. One hundred and twenty consecutive patients with active brucellosis were enrolled. The commonest way of transmission was ingestion of milk products from diseased animals. Brucella melitensis was isolated in the specimens of 31 (45.5%) of 68 patients. The commonest abnormalities on physical examination were fever (66.6%), hepatomegaly (63.3%) and splenomegaly (56.6%). Osteoarticular involvement was found in 34 patients (28.3%). Fifteen (12.5%) patients had ocular involvement. Hepatitis, orchiepididymitis, pulmonary involvement and meningitis were found in one (0.8%), four (6.8%), three (2.5%) and one (0.8) patient, respectively. The commonest haematological abnormalities were relative lymphomonocytosis (71.6%) and anaemia (36.6%). In conclusion, brucellosis continues to be a common health problem in communities where the consumption of unpasteurised dairy products is common. Since prevention is as important as early diagnosis in reducing the morbidity of brucellosis, we suggest that improving current health policies with additional educational programmes is essential.

Abstract: A retrospective postal survey of 21 000 medical laboratory workers in England and Wales showed 18 new cases of pulmonary tuberculosis in 1971, a five-times increased risk of acquiring the disease compared with the general population. Technicians were at greatest risk, especially if they worked in morbid anatomy departments. Of the 35 cases of hepatitis, the technicians were again the occupational group most likely to acquire the disease. Microbiology staff were twice as likely to report shigellosis as those in other pathology divisions but only one case of brucellosis was reported in the whole laboratory population. A similar survey carried out in 1973 of 3000 Scottish medical laboratory workers corroborates the results from England and Wales. Medical laboratory workers continue to experience a considerable risk of developing an occupationally acquired infection. Improvements in staff safety and health care seem to be necessary.

Abstract: The structure of the lipid A from Rhizobium etli and Rhizobium leguminosarum lipopolysaccharides (LPSs) lacks phosphate and contains a galacturonosyl residue at its 4' position, an acylated 2-aminogluconate in place of the proximal glucosamine, and a very long chain omega-1 hydroxy fatty acid, 27-hydroxyoctacosanoic acid (27OHC28:0). The 27OHC28:0 moiety is common in lipid A's among members of the Rhizobiaceae and also among a number of the facultative intracellular pathogens that form chronic infections, e.g., Brucella abortus, Bartonella henselae, and Legionella pneumophila. In this paper, a mutant of R. leguminosarum was created by placing a kanamycin resistance cassette within acpXL, the gene which encodes the acyl carrier protein for 27OHC28:0. The result was an LPS containing a tetraacylated lipid A lacking 27OHC28:0. A small amount of the mutant lipid A may contain an added palmitic acid residue. The mutant is sensitive to changes in osmolarity and an increase in acidity, growth conditions that likely occur in the nodule microenvironment. In spite of the probably hostile microenvironment of the nodule, the acpXL mutant is still able to form nitrogen-fixing root nodules even though the appearance and development of nodules are delayed. Therefore, it is possible that the acpXL mutant has a host-inducible mechanism which enables it to adapt to these physiological changes.

Abstract: In this study we aimed to determine if Campylobacter had the ability to induce an adaptive tolerance response (ATR) to acid and/or aerobic conditions. Campylobacter jejuni CI 120 was grown to the appropriate phase in Brucella broth under microaerobic conditions. Cells were initially adapted to a mild stress (pH 5.5) for 5 h prior to challenge at pH 4.5, a lethal pH. Survival was examined by determining the numbers of viable cells on Campylobacter blood free selective agar base. Stationary phase cells adapted at pH 5.5 induced an ATR that enabled a 100-fold greater survival compared to an uninduced culture. Aerobic adaptation also protected the cells against acid challenge. The cross protection provided a 500-fold increase in survival when compared to unadapted cells. The incorporation of chloramphenicol during the induction period eliminated the ATR and resulted in death kinetics similar to an uninduced culture. These data suggest that Campylobacter spp. have the ability to induce an ATR to sublethal treatments, which increased their ability to withstand subsequent stresses.

Abstract: Protection against brucellosis involves both cellular and humoral effectors not yet fully appreciated. Living or killed vaccines can protect against the infection itself or only against abortion. For official controls, vaccines (or new procedures of vaccination) must first be characterized pharmacologically and tested for innocuity. Protection must be tested on natural hosts with a reference vaccine (S19 or Rev. 1) by the agreed method which reproduces the natural infection and measures immunity in toto. Control and vaccinated females are challenged by the conjunctival route at mid-pregnancy under standard conditions (strain, dose) to measure the resulting infection by bacteriological analysis of excretion at parturition and of infection in target organs at slaughter. Results are principally expressed by the infection rate which should be +/- 95% in the control group. In the new vaccine group the rate should be equivalent to, or lower than, the reference vaccine group. To be statistically valid, at least 30 animals per group are required. For routine controls, laboratory models using guinea pigs, not well standardized, inaccurate and expensive, have long been proposed. The mouse model, extensively studied and standardized, should now be preferred to the guinea pig model. In the mouse model, residual virulence of a living vaccine is estimated by the time required by 50% of the mice to eradicate the strain from their spleen (Recovery Time 50%). Immunogenicity is measured by the ability of mice to restrict their splenic infection after a virulent i.p. challenge at a dose (B. abortus 544; 2 x 10(5) cfu) chosen in order that all mice were still infected 15 days post challenge.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: One hundred and sixty-six presumed brucellosis patients were included in the study. These patients were classified as primary (91), relapse (18) and suspected (57) cases according to their clinical presentations, and serologic and microbiologic test results. Primary and relapse cases were evaluated retrospectively according to age, sex, residence, routes of transmission, clinical and laboratory findings, treatment regimens, duration of treatment, and relapse rates. Of the 109 primary and relapse patients, 57 were male and 52 female. The ages of the patients ranged between 16-75 (mean age 40.2). The percentages of the urban and rural residence of the patients were 41.3% and 58.7%, respectively. The most common mode of transmission was consumption of unpasteurized milk and milk products (67.9%). Malaise, fever and sweating were the most frequently observed symptoms (96.3%, 95.4%, 91.7%, respectively). The most common signs were fever (97.2%), splenomegaly (59.6%), and hepatomegaly (37.6%). The liver was the most frequently involved organ (21.1%). Almost all (99.1%) patients were serologically positive. However, the positivity rate of culture was low (15.6%). The most frequently preferred antimicrobial regimen was rifampin and doxycycline combination. The relapse rate was 8.3%. Brucellosis is still prevalent in Turkey as in many other countries in the Mediterranean basin. The clinical presentation of the disease may show regional variations. Patients with a history of occupational or nutritional contact with the bacterium and with a compatible clinical picture should be examined using appropriate diagnostic techniques before any attempt to prescribe an antimicrobial.

Abstract: Eleven brucellosis cases were identified in three municipalities of Cordoba (Andalucia, Spain). A case-control study was conducted, selecting three cases per control. Persons having eaten unpasteurized raw goat cheese produced in a farmhouse located in the epidemic territory, were at higher risk for presenting brucellosis (OR=21.6, IC95%=1.6-639.8). Brucella melitensis serovar 3 was identified in clinical specimens and in goat tissue and milk samples from the herd's farmhouse. Preventive measures were implemented and the outbreak was stopped after the withdrawal of all suspicious cheeses from the market, additional sanitation of the farmhouse and health promotion activities.

Abstract: An investigation of dangerous bacterial pathogens was conducted to determine the usefulness of automated rRNA operon ribotyping (RiboPrinter system) to identify species. A total of 26 isolates comprising Bacillus anthracis, Brucella spp., Burkholderia mallei, Francisella tularensis, and Yersinia pestis were tested using restriction endonucleases EcoRI, PstI, PvuII and AseI. The main problem was that the system's database-relying on EcoRI as restriction enzyme-does not contain the essential dangerous pathogens. B. anthracis was misidentified as B. cereus and Y. pestis as Y. pseudotuberculosis. Two isolates of F. tularensis ssp. holarctica were falsely identified as Vibrio cholerae. This study underscores that riboprint patterns generated with a single restriction enzyme are not always unique for each of the species tested. Using more than one enzyme, the RiboPrinter proved to be a valuable primary typing method. Databases of commercially available systems for the identification of bacteria should include the most important dangerous pathogens.

Abstract: AIM: To study stress resistance mechanisms in Campylobacter spp. METHODS AND RESULTS: Campylobacter strains were grown to the appropriate phase in Brucella broth. The cells were diluted into either cell-free spent medium (obtained by filtration of a grown culture) or a freshly prepared medium and the pH reduced to 4.5, a lethal pH value. At suitable time intervals survivors were enumerated on Campylobacter blood free selective agar base. The cell-free spent medium from mid-exponential and stationary phase had a protective effect on acid and thermal stress in Campylobacter jejuni CI 120, a natural isolate. The protective effect of the extracellular compound was not significantly inactivated by boiling, but was inactivated by proteinase. CONCLUSIONS: The present study suggests that a protein (or proteins) accumulated by C. jejuni CI 120 during growth may play an active role in the induction of stress responses and that this protein is heat stable. SIGNIFICANCE AND IMPACT OF THE STUDY: The results indicate that C. jejuni CI 120, a natural isolate, has the ability to use extracellular signalling mechanisms to induce tolerance to stress factors. This is a major advancement in the understanding of the physiological basis for survival of C. jejuni in the environment.

Abstract: In Somalia, one of the world's largest dromedary populations of about 5.3 million animals are kept by nomadic pastoralists under traditional management. Interest in the development potential of camel herds in the semi-arid areas of central Somalia initiated an investigation to determine the productivity of herds, their major diseases and likely associations among these parameters. Using a systems approach, data were collected for herd production parameters, environmental factors, management and production systems, and health variables. One thousand and thirty nine camels in 33 herds were studied in the central regions of Somalia. Trypanosoma evansi prevalence ranged from 1.7% in blood-smears to 56.4% using enzyme-linked immunosorbent micro-assay (microELISA). Seroprevalence for brucellosis was determined as 1.9% by the standard agglutination test (SAT) and 0.3% by the complement fixation test (CFT). Using multiple regression, 15% of the total variation of the general fertility rate was explained by the results of the microhaematocrit centrifugation technique (MHCT) and the microELISA for T. evansi, CFT results for brucellosis, herdsize, and young stock death rate. Among herd production variables, herd size differed significantly for different management units. Young stock death rates, as well as general fertility rates varied in the ecological subzones with a marked effect in the zones labeled "Inland". Various other associations were noted among demographic, husbandry and disease variables. The importance of trypanosomosis and brucellosis to the productivity of herds and measures to control their limiting effects on production were discussed.

Abstract: Campylobacter jejuni has been researched in raw milk, skin and excrement of chicken. All milk samples are negatives but 66% of chicken skin and 12% of chicken excrements are positives. An enrichment phase using brucella broth added with horse blood and antibiotics is necessary.

Abstract: One of the advantages of ELISA techniques is to achieve a higher degree of safety by standardization. Bases of these standardizations should be, if available, international or national standards in combination with cutoff definitions. The cutoff definitions should be expressed, separate for the main kinds of samples, defined as a dilution of the standard. Those definitions can replace the mostly awkward and involved test descriptions as usual in official regulations. To guarantee a high grade of reliability it is a matter of urgent necessary to test (if possible by the producer) each serial (lot) of ELISA and to describe their capacities with regard of sensitivity and specificity. Separate pretests may be done with the main kinds of samples with well known reactions (negative and weak positive). The results are to compare to those of the cutoff-related dilutions of the standard in the same matrix. These descriptions of the capacities of each serial (lot) are safe bases for use of the serial. Key concepts are the quotients of the minimum positive value divided by the maximum negative or alternatively the mean value of the positive minus the threefold standard deviation divided by the mean value of the negative plus the threefold standard deviation.

. THE VALUE of the milk ring test (M.R.T.) and related tests in the detection of brucella organisms in milk supplies. Monthly bulletin of the Ministry of Health and the Public Health Laboratory Service. 1956 May; 15; 85-95. [PubMed: 13321874].

GUERRA M. [Research on the survival and the persistence of various bacteriological characteristics and on the pathogenicity of Brucella in cream.]. Bollettino dell'Istituto sieroterapico milanese. 1957 May-Jun; 36(5-6); 306-10. [PubMed: 13460017].

GEORGE JT, PAYNE DJ. Tuberculosis from T.T. milk, with a note on the frequency of Brucella abortus in consumer milk. Monthly bulletin of the Ministry of Health and the Public Health Laboratory Service. 1961 May; 20; 99-102. [PubMed: 13704453].

Abstract: Liposomes were evaluated for their effectiveness as vaccine carriers in the potentiation of the mouse humoral response to the lipopolysaccharide (LPS) and O-polysaccharide (OPS) antigens of Brucella abortus. LPS and OPS were extracted from a pathogenic strain of B. abortus and were encapsulated within multilamellar vesicles. Groups of mice, immunized with liposome-encapsulated and free LPS or OPS, were bled weekly and the specific IgM and IgG levels in the sera were determined by an indirect fluorogenic enzyme-linked immunosorbent assay. Humoral response to these antigens were found to be dose-dependent. Mice immunized with LPS and OPS encapsulated within liposomes were found to have significantly higher IgG levels than mice immunized with free LPS and OPS. In addition, the antibody levels in mice that were immunized with liposome-encapsulated LPS and OPS were more sustained and remained at elevated levels--even after 5 weeks post immunization. As expected, OPS was found to be less immunogenic than LPS, but multiple injections of OPS encapsulated within liposomes greatly improved the immunogenicity. These results indicate that the humoral response to LPS and OPS of B. abortus can be enhanced when these antigens are encapsulated within liposomes.

Abstract: Intravenous injection of large numbers of live, virulent Brucella abortus in chickens resulted in the appearance in the serum of a viral inhibitor indistinguishable from interferon. Inhibitor was detected as early as 3 hours after inoculation of brucellae and reached a peak between 6 and 12 hours.

MATHUR TN. THE ROLE OF THE GOAT IN HUMAN BRUCELLOSIS IN INDIA WITH PARTICULAR REFERENCE TO INFECTION WITH BRUCELLA MELITENSIS. The Journal of the Association of Physicians of India. 1964 Dec; 12; 805-13. [PubMed: 14253378].

Cooper CW. Risk factors in transmission of brucellosis from animals to humans in Saudi Arabia. Transactions of the Royal Society of Tropical Medicine and Hygiene. 1992 Mar-Apr; 86(2); 206-9. [PubMed: 1440791].

Abstract: A case-control study was undertaken to determine for the first time the specific aetiology of animal to human transmission of brucellosis in Saudi Arabia. Cases consisted of all patients with brucellosis attending the primary care clinic of the Riyadh Al-Kharj Hospital programme in central Saudi Arabia. A sample of individually matched controls was selected concurrently from patients attending the same clinic for unrelated problems. Cases and controls responded to a 48 item questionnaire on exposure to established risk factors for brucellosis. Greatest risk was found to be associated with indirect contact with animals (the consumption of unpasteurized dairy products), as opposed to direct contact with animals. When specific animal products were considered, greatest risk was associated with the consumption of milk and laban (buttermilk), as opposed to cheese or uncooked liver. When specific animal species were considered, greatest risk was associated with products derived from sheep and goats as opposed to camels and cattle. When direct contact with animals was considered, the study found a very high risk associated with assisting in animal parturition, but no significant risk associated with other direct (unspecified) animal contact.

Abstract: Using the methods of acid platelet agglutination, agglutination, agglutination with 2-mercaptoethanol, Coombs' reaction, and complement binding reaction, 152 serum specimens taken from patients with diagnosed brucellosis, were subjected to studies. The following per cent of seropositive results was shown: acid platelet agglutination--11.2%, agglutination--40.1%, agglutination with 2-mercaptoethanol--34.4%, Coombs' reaction--42.8%, complement binding reaction--7.9%. The acid platelet agglutination reaction cannot be used for screening for brucellosis in humans.

Abstract: 1. Our study showed a prevalence of 5.33% in animal handlers working in an urban city like Pune. The prevalence would definitely be higher in a population from a rural area. 2. All these cases who showed presence of antibodies to B. abortus, had varied clinical manifestations, characteristic of the protean manifestations in brucellosis. Likewise in our study we had cases ranging from arthritis, abortions and genito urinary manifestations. 3. All the antibody positive cases had significant antibody titres. The clinicians miss many cases of brucellosis because it is not considered as an alternative diagnosis. The clinician should keep in mind the possibility of an occupational or environmental exposure in cases of P.U.O. It would also be worthwhile to create awareness of the disease in such professions so that necessary precautions and periodic screening of such occupationally exposed people can be done. Studies are needed to assess the role of brucellosis as a cause of morbidity in India, which had not received the attention it deserved. Prevention of human brucellosis focuses mainly on elimination of infection in cattle along with hygiene, vaccine, and effective heating and pasteurization of dairy products and related foods.

Abstract: Beef and dairy cattle serum samples, collected during 2000 at sale barns throughout Georgia, were obtained from the Georgia State Brucellosis Laboratory and were used to conduct a retrospective epidemiological study. Statistical samplings of 5,307 sera, from over 200,000 sera, were tested for antibodies to Mycobacterium avium ssp. paratuberculosis, (Johne's disease) using a commercial enzyme-linked immunosorbent assay test kit. An overall period seroprevalence in all classes of cattle tested was 4.73%. The period seroprevalence in dairy cattle was 9.58%, in beef cattle it was 3.95%, and in cattle of unknown breed it was 4.72%. It was concluded that the seroprevalence of Johne's disease in cull beef and dairy cattle in Georgia is economically significant.

Abstract: Three dogs reared on a dairy farm with a high incidence for Brucella abortus were serologically positive for B. abortus and no other Brucella spp. The identity of the organism was confirmed to be B. abortus by AMOS (abortus melitensis ovis suis)-polymerase chain reaction with specific primers for B. canis. One hundred percent homology of the canine isolate and the bovine pathogen isolated from the farm was demonstrated. The only possible source of infection was infected cattle on the same farm. It is suggested that dogs be routinely included in brucellosis surveillance and eradication programs.

Abstract: As a part of a research-and-action partnership between public health and veterinary medicine, the relationships between the seroprevalences of brucellosis and Q-fever in humans and livestock were evaluated in three nomadic communities of Chad (Fulani cattle breeders, and Arab camel and cattle breeders). Nomad camps were visited between April 1999 and April 2000. A total of 860 human and 1637 animal sera were tested for antibodies against Brucella spp., and 368 human and 613 animal sera for Coxiella burnetii. The same indirect ELISA was used for livestock and human sera, and the test characteristics for its use on human sera were evaluated. Twenty-eight people were seropositive for brucellosis (seroprevalence 3.8%). Brucella seroprevalence was higher in cattle (7%) than other livestock, and brucellosis seropositivity was a significant factor for abortion in cattle (OR=2.8). No correlation was found between human brucellosis serostatus and camp proportions of seropositive animals. Q-fever-seropositive blood samples were taken from 11 Arab camel and 4 Arab cattle breeders (seroprevalence 1%). Being a camel breeder was associated with Q-fever seropositivity in humans (OR=9). Camels had the highest Q-fever seroprevalence (80%) among livestock species. Although there was high-risk human behaviour for the acquisition of brucellosis and Q-fever from livestock through raw-milk consumption (98%) and contact with placentas of livestock (62%), we concluded that seroprevalences in humans were relatively low (likely due to limited active foci in livestock).

Abstract: AIM: The aim of the present experiment was to study the effect of different chlorhexidine regimens on the number of bacteria in saliva, and on de novo plaque formation. MATERIAL AND METHODS: Ten subjects with gingivitis, but no signs of destructive periodontitis, were recruited. Following a screening examination, the volunteers were given oral hygiene instruction, meticulous scaling and professional mechanical tooth cleaning (PTC). The PTC was repeated once every 3 days during a 2-week period to establish healthy gingival conditions. The study was designed as a double-blind cross-over clinical trial including three phases. Each experimental phase comprised one preparatory period of 7 days and one plaque accumulation period (no oral hygiene measures) of 4 days. During all preparatory periods, the volunteers (i) performed mechanical tooth cleaning using a toothbrush and dentifrice and (ii) were, in addition, given two sessions of PTC. The final PTC was delivered after bacterial sampling had been made on Day 0. Preparatory period A: the participants continued the self-performed plaque control regimen that employed only mechanical means. Preparatory period B: the participants were in addition instructed to rinse and gargle, twice daily, with a 0.2% chlorhexidine mouthrinse. Preparatory period C: in addition to the above, the participants were instructed to brush the dorsum of the tongue for 60 s, twice daily, with a 1.0% chlorhexidine gel. Following each plaque accumulation period, there was a 10-day washout interval. The presence and amount of dental plaque (QHI) was scored after 1, 2 and 4 days of no oral hygiene. Samples of saliva were obtained on Day 0 and after 1 and 2 days. The samples were placed on Brucella agar plates and incubated (anaerobically) for 5 days. The total number of colony-forming units was determined and used to estimate the density of bacteria in saliva. RESULTS: In period A, the mean QHI increased from 1.0 (Day 1) to 1.4 (Day 2) and 2.1 (Day 4). The corresponding scores for periods B and C were 0.5, 0.8, 1.6 and 0.3, 0.8, 1.2, respectively. At all re-examination intervals more plaque formed during period A than during periods B and C. Further, during period C, less plaque formed than that during period B. Saliva samples from Day 0 in period A contained a larger number of TVC than the baseline samples in periods B and C. There was no significant difference in TVC among the groups on Day 2. CONCLUSION: The daily use of chlorhexidine as an adjunct to mechanical tooth cleaning markedly reduced the number of microorganisms that could be detected in saliva. The number of salivary bacteria may have influenced the amount of plaque that formed during an early phase of no oral hygiene.

Abstract: Sera from 2,630 apparently normal adult camels (Camelus dromedarius) raised in central Saudi Arabia (Riyadh and Al-Kharj cities) were examined serologically by the Rose Bengal and standard United States of America Brucella plate agglutination tests. The overall seroprevalence of brucellosis in the restricted populations of tested camels was 8%. The seroprevalence of brucellosis among camels raised in small numbers in the backyards of 24 houses in Riyadh and those intensively raised on one large camel farm near Al-Kharj were 4.3% and 8.6% respectively. Fresh milk samples from 100 brucellosis seropositive camels from Riyadh and Al-Kharj were cultured on Brucella-selective media. Brucella melitensis biovars 1 and 2 were isolated and identified from 26 camels. Epidemiologically, brucellosis in camels in central Saudi Arabia appeared to be connected with B. melitensis infection of sheep and goats, and also represents a serious public health risk.

Abstract: We report herein a case of Bartholin's gland abscess caused by Brucella melitensis. Clinical microbiology laboratory workers in areas where this disease is endemic should be familiar with the bacteriological features of this organism and consider the possibility of a brucellar etiology in a broad range of clinical settings.

Abstract: Brucellosis is a zoonotic disease of worldwide distribution that mainly affects persons working with domestic animals and animal products. Despite being controlled in many developed countries, the disease remains endemic in many parts of the world, including Latin America, the Middle East, Spain, parts of Africa, and western Asia. The disease is mainly transmitted to humans through the ingestion of raw milk or non-pasteurized cheese contaminated with one of the four Brucella species pathogenic to humans. The clinical presentation can vary from asymptomatic infection with seroconversion to a full-blown clinical picture of fever, night sweats and joint manifestations; rarely, there is hepatic, cardiac, ocular or central nervous system involvement. Since travelers may be affected, travel health physicians need to know the clinical presentation of patients with brucellosis and preventive strategies.

Abstract: To perform a comparative evaluation of the indirect enzyme-linked immunosorbent assay (iELISA) in milk, for the detection of Brucella abortus in cattle, 1,523 milk samples were taken from individual animals and bulk milk belonging to 200 herds in the province of Cundinamarca, Colombia. All these herds were part of the official programme of monitoring free herds and determining herd prevalence in the province. The samples were submitted simultaneously to the milk ring test and the iELISA. A total of 584 individual and 497 bulk milk samples taken from free herds were considered negative, and served to determine the initial ELISA cut-off point. An optimised determination of the cut-off point involved an operational analysis, using a statistical programme. In the analysis, the total population was considered: 756 individual positive and negative samples, as defined by herd history and previously established serologies. The cut-off point was set at > 0.20 optical density units, which, expressed in percentage positivity, corresponded to 20.5% against the controls, with a sensitivity of 95.3%, a specificity of 95.1%, and a confidence interval of 95%. Indirect confirmation of the state of infection was made with competitive ELISA in the serum of the individual animals considered positive, and an attempt was made to determine bacterial presence by isolation in culture and by reverse transcription-polymerase chain reaction. The results confirm that the indirect ELISA is highly sensitive and specific, provides confirmed results in strict quality-control conditions, and may be used to test a large number of herd or individual samples, thus enhancing the efficiency of surveillance programmes and control campaigns.

Abstract: Brucella is responsible for one of the major worldwide zoonoses. Over the last century, several vaccines have been used against brucellosis. Among these, the rough vaccine Brucella abortus RB51 was introduced with the idea that it would not interfere with the diagnosis of brucellosis. Recently, RB51 has been isolated from milk and vaginal exudates from vaccinated cows, thus raising the possibility of extensive bacterial replication in these animals. We hypothesized that shedding of RB51 might be related to a change in its intracellular cell cycle. Therefore, we have compared the intracellular trafficking in CHO cells of the virulent B. abortus 2308 and two RB51 strains, the vaccinal strain and the one isolated from cow milk. Both RB51 strains were transiently observed in phagosomes characterized by the presence of the early endosomal marker EEA1 and then were found in cathepsin D-enriched lysosomal compartments, in which they eventually underwent degradation at later post-infection times. In contrast, the virulent 2308 strain replicated within the endoplasmic reticulum. These results suggest that a change in intracellular trafficking cannot account for Brucella shedding in adult vaccinated cows.

Abstract: Great Britain (GB) has been "Officially Brucellosis Free" (OBF) since 1991; because this disease has both public-health and international-trade implications, it is in the country's interest to maintain this freedom. A quantitative risk-assessment model was developed to determine the annual risk of importing brucellosis-infected breeding cattle into GB from Northern Ireland and the Republic of Ireland. (These countries exported the largest number of cattle into GB and were not brucellosis free during the development of the assessment in 2000.) We predicted that we can expect to import brucellosis from Northern Ireland every 2.63 years (1.89, 4.17) and from the Republic of Ireland, every 3.23 years (2.13, 5.88). The estimates of risk are sensitive to the assumed proportion of animals missed during routine surveillance that originate from OBF herds and the uncertainty associated with the surveillance test sensitivities. As a result of the assessment, the Department for Environment, Food and Rural Affairs (Defra) introduced post-calving testing for all cattle imported into British herds.

Abstract: OBJECTIVES: The intracellular antibiotic efficiency of gentamicin-loaded microspheres in the context of Brucella-infected murine monocytes was examined in vitro with a view to developing improved therapies for the treatment of brucellosis. METHODS: Biodegradable microspheres made of end-group capped and uncapped poly(lactide-co-glycolide) 50:50 (PLGA 50:50 and PLGA 50:50H) and containing gentamicin sulphate were used to target Brucella abortus-infected J774 monocyte-macrophages. The infected cells were treated with 15 micro g of free or microencapsulated gentamicin and the efficacy of the treatments was measured after 24 h. RESULTS: The particle sizes were below 8 micro m and in vitro release of gentamicin from the microspheres followed a continuous (PLGA 50:50H) or a multiphasic (PLGA 50:50) pattern over 50 days. Treatment with gentamicin microencapsulated into the end-group uncapped PLGA 50:50H microspheres, decreased significantly the number of intracellular bacteria (typically by 2 log(10)) in comparison with untreated infected cells. Addition of 2% poloxamer 188 to the microsphere dispersion medium further reduced the infection (3.5 log(10)). Opsonization of the particles with non-immune mouse serum had no effect on the antibacterial efficacy of the microspheres. End-group capped PLGA 50:50 type microspheres containing the antibiotic were less effective at reducing intracellular bacteria ( approximately 1 log(10) reduction), although addition of poloxamer 188 to the dispersion medium again enhanced their intracellular antibacterial activity. Placebo PLGA 50:50 and PLGA 50:50H microspheres had no bactericidal activity. CONCLUSIONS: The results indicate that PLGA 50:50-microencapsulated gentamicin sulphate may be suitable for efficient drug targeting and delivery to reduce intracellular Brucella infections.

Abstract: Chickens injected subcutaneously at one day of age with oil-in-water emulsions of the nonioinic block polymer T150R1 developed higher titres to sheep erythrocytes and killed Brucella abortus than did untreated chickens when they were immunised with the antigens one and two weeks later. The increase was IgM only. Cell-mediated immunity was not affected as measured by blastogenic responses. Morphometric analysis of tissue sections of bursae of Fabricius suggested the polymers were stimulating more rapid migration of B cells to peripheral organs.

Abstract: An alloantigen is a genetically determined cell-surface molecule detected by specific antisera. An identifying letter has been assigned to each genetic locus responsible for the 12 distinct families of alloantigens: A, B, C, D, E, H, I, J, K, L, P, and R. The genes of each system segregate independently of the other systems, except that the A and E are very closely linked (0.5 centimorgans). Selection experiments over numerous generations have revealed distinct changes in gene frequency of the A-E alloantigens, suggesting immune responses associated with susceptibility to coccidiosis, response to immunizations with SRBC, and selection for size of the bursa of Fabricius. Immune response effects of the C system of alloantigen genes are indicated by distinct gene frequency changes following selection for response to SRBC, selection for size of bursa of Fabricius, and macrophage nitrite production after lipopolysaccharide (LPS) stimulation. Immune response effects of the D system of antigens are indicated by data from genetic selection for response to immunization with SRBC, selection for bursa size, and macrophage nitrite and cytokine interleukin (IL)-6 production following LPS stimulation. Immune response effects of the I system genes are indicated by distinct gene frequency changes in lines selected for bursa size and within family comparisons for macrophage nitrite and cytokine IL-6 production following LPS stimulation. Effects of the L system, consisting of only 2 alleles, are indicated by the gene frequency changes following selection for bursa size, direct comparison of genotypes within families for monocyte phagocytosis, susceptibility to coccidiosis, outcome of Rous sarcomas, and immune responses to SRBC and Brucella abortus. Genotypes of the P alloantigen system were directly compared within families of fully pedigreed chicks with significant differences for monocyte phagocytosis. An experimental procedure for simultaneously testing for immune responses of genotypes of 9 of the alloantigen systems (A, B, C, D, E, H, I, L, and P) has been established by producing test progeny from a single cross of parent lines segregating for genes of each of the systems.

Abstract: STUDY DESIGN: A case report. OBJECTIVES: To present and discuss some of the difficulties in the diagnosis of brucellar spondylitis. SETTING: Ankara University, Ibni Sina Hospital, Turkey. METHODS: We report a patient with paraplegia, misdiagnosed as having a malignancy or tuberculosis who actually suffered from brucellar spondylitis. Diagnosis was established by her history and a compatible clinical picture together with a standard tube agglutination (Wright test) titer of > or =1/160 of antibodies for brucellosis. The patient was treated with oral doxycycline, rifampicin, and ciprofloxacin combination. RESULTS: At the end of the treatment, the blood Brucella Wright and anti-human globulin T titer levels decreased. Her lower limb weakness improved. She could walk, and climb stairs with the help of a cane. Urinary retention and fecal incontinence also resolved. CONCLUSION: Brucellosis is a systemic infection involving the musculoskeletal and nervous systems. Spondylitis frequently occurs in elderly patients. An early diagnosis of brucellar spondylitis can often be difficult. In endemic regions, as in the case of our country, brucellar spondylitis should always be considered in the differential diagnosis of older patients with back pain and constitutional symptoms. An early diagnosis will help to prevent the development of more severe complications such as spinal cord compression.

Abstract: This work evaluates the influence of different pharmaceutical auxiliaries (Pluronic F68, polyvinylpyrrolidone [PVP] or Tween 20), when mixed with an antigenic extract from Brucella ovis (hot saline; HS), on the characteristics of the resulting poly(epsilon-caprolactone) (PEC) and poly(lactide-co-glycolide) (PLGA) microparticles. In all cases, PEC microparticles were smaller than PLGA ones. Concerning the HS loading, PLGA microparticles were highly dependent on the type of the excipient used, whereas all the PEC formulations displayed similar encapsulation efficiencies. For both types of microparticles, the presence of PVP induced a burst release effect. On the contrary, the use of Tween 20 or Pluronic F68 dramatically modified this profile. For PLGA-Tween 20 and PEC-Pluronic F68 microparticles, the HS was released in a pulsatil way during the first 7 days followed by a continuous release for at least 3 weeks. The antigenicity of the HS components was kept in all cases. Phagocytosis by murine monocytes showed a clear difference based just on the hydrophobicity of the polymer, being PEC microparticles better engulfed. Cell activation quantified by the release of H2O2 did not showed major differences between batches, however, microparticles of PEC and Pluronic F68 induced the highest nitric oxide production. Together, these results confirm the advantageous qualities of the "HS-PEC-Pluronic F68 microparticles" as favorable candidate for vaccine purposes against brucellosis.

Abstract: A study was conducted to estimate the seroprevalence of Brucella ovis infection in rams in the Estrie and Bas-Saint-Laurent regions (Quebec). Rams sera (n = 258) were serologically evaluated from 224 rams in 30 commercial flocks and from 34 rams at 2 slaughterhouses by using an enzyme linked immunosorbent assay. Epididymides and testes were examined by palpation on farms and microscopically for culled rams. No ram was seropositive to Brucella ovis or had lesions suggestive of brucellosis from the farm or slaughterhouse surveys.

Abstract: We report two cases of brucellosis in members of a hunt club, both of whom had killed and dressed wild boars. Serologic studies usually suffice for screening patients with suspected or possible brucellosis; however, and as illustrated by our index case, one should ask the laboratory to dilute serum out beyond the customary 1:160 titer if results are negative yet the clinical suspicion high. Hunters should be advised to wear gloves prior to dressing wild mammals.

Abstract: Isolated reports of brucellosis among family members have been documented. The aim of this study is to determine if active serological screening of the households' members of acute brucellosis cases will detect additional unrecognized cases. From May 2000 to October 2001, patients with acute brucellosis were enrolled and their household members were serologically screened for brucellosis using the Standard Agglutination Test (SAT). Fifty-five index cases with acute brucellosis and 404 household members were enrolled. The majority of index cases (48%) were young adults, and 79% were illiterate. Ownership of animals and ingestion of unpasteurized raw milk were reported by 45 and 75% of the index cases respectively. Of the 55 families screened, 23 (42%) had two family members or more with serological evidence of brucellosis and 32 (58%) had only the index case. Households of > or = 5 members and a history of raw-milk ingestion by family members were risk factors associated with the seropositives (P < 0.05). Of the 404 household members screened, 53 (13%) were seropositive; of these 39 (74%) were symptomatic, and 9 (35%) had brucella bacteraemia. Symptomatic seropositives tended to have bacteraemia and higher brucella antibody titres compared to asymptomatic seropositives (P < or = 0.05). Screening family members of an index case of acute brucellosis will detect additional cases.

Abstract: The communal grazing system is generally understood to have a low input, low output type of management. However, the actual inputs and outputs of the farmers are not well known and the farmers are often unaware of their problems. Although the causes of low calving percentage are well understood in commercial beef farming enterprises in South Africa, the same is not true for communal farming systems. The aim of this study was to determine the reproductive performance of beef cattle on a communal farming system in Jericho, North West Province. Ten farmers from five villages with a total of 265 cows and 13 bulls were purposively selected. The selection criteria were that each farmer had to have a minimum of 10 breeding cows and a bull and be willing to participate in the study. This was followed by a 12-month longitudinal study with monthly herd visits where cows were examined rectally and bulls (n = 13) were subjected to a single breeding soundness evaluation. The calving percentage was found to be 37.7%. This is lower than the recorded percentages for commercial beef cattle on extensive grazing. The factors playing a role in low calving percentage were ranked using field data. From this it appeared that failure of cows to become pregnant was the main cause of poor calving percentage as opposed of loss of calves through abortion or resorption. Sub-fertility of the bulls was found to be of great significance and it is proposed that this be included in extension messages and that bulls be fertility tested routinely. Poor body condition score of cows, mainly caused by poor management, was also considered to play a major role in reducing pregnancy rates. Infectious diseases like trichomonosis, campylobacteriosis and brucellosis played a much leser role than anticipated.

Abstract: A total of 5726 blood specimens (from children aged 14 years and younger) were studied for the serological evidence of brucellosis. Ninety-three (1.6 per cent) showed diagnostic agglutinin titres with a geometric mean titre of 403 (SD +/- 547). Forty-three (59.7 per cent) blood specimens yielded the growth of Brucella melitensis. Thirty-nine patients (41.93 per cent) were shepherds, who constituted the major occupational group affected in the present series. More than 60 per cent of the patients had a history of both consumption of fresh goat's milk and close animal contact. The habit of consuming fresh goat's milk to obtain relief from chronic ailments was noted in nine patients. Seventy-three (78.49 per cent) were males and 20 (21.51 per cent) were females, with a male to female ratio of 3:1. The disease occurred mainly in the school age group (mean age 10.3 years). All the patients had an acute history of less than 2 months. Forty-nine (52.68 per cent) patients presented with persistent fever, 19 (20.43 per cent) with joint pain, and the rest with a combination of fever and joint pain with and without low backache, fever being the commonest complaint. One case presented with involuntary movements of limbs alone and the other with burning feet only. Pityriasis alba was the consistent physical finding, with fever in the majority of the patients. The major joint found to be involved was the knee (52.77 per cent). The synovial fluid obtained from the knee joint of five patients demonstrated Brucella agglutinins and also three grew B. melitensis. Eight patients presented with complications that included skin lesions (3), carditis (2), neurobrucellosis such as chorea (1), peripheral neuritis (1), and meningitis (1). Brucella melitensis biotype 1 was successfully isolated from the papular eruption of one out of three cases who presented with skin lesions. To our knowledge this is the fourth confirmed isolation of B. melitensis from skin lesions with brucellosis, reported in the literature. The cerebrospinal fluid obtained from the meningitis patient was positive for B. agglutinins. To our knowledge chorea of brucellar origin appears to be the first case reported in the literature. In 15 cases (16.13 per cent) brucellosis was suspected clinically whereas 78 (83.87 per cent) cases, only serological evidence of brucellosis confirmed the diagnosis. None of the cases relapsed. In our experience an initial combination therapy with a three-drug regimen followed by a two-drug regimen for a minimum of 6 weeks has been found to be effective in the prevention of a relapse.

Abstract: Brucellosis is a zoonotic disease that is encountered in sheep rather frequently. In this study, 100 sheep diagnosed with brucellosis that had aborts and 40 healthy sheep were used as materials. Analyses for Cu, Zn, Fe, Cr, Ca, Mg, and K were performed with the atomic absorption spectrophotometric method on blood collected from vena jugularis of all the sheep and rheumatoid factor levels were determined by the nephelometry method. Although it was found that Cu, Ca, and rheumatoid factor values in the sera of the sheep with brucellosis were high when compared to the control group (p<0.001, p<0.05, p<0.001, respectively), their serum Zn values were low (p<0.05). No significant changes in serum Cr, Fe, K, and Mg levels were found.

Abstract: The organisation of veterinary public health (VPH) services in countries of Latin America and the Caribbean has resulted from an historical process which has consolidated the structure of these services and the concept of VPH. This concept comprises five principal plans of action: promotion of animal health in order to increase the production of protein of animal origin; protection of foods for human consumption; vigilance, prevention, control and eradication of zoonoses; promotion of environmental protection, together with the development of biomedical models. These plans of action serve the purpose of improving human health and well-being, which is essential for the socioeconomic development of populations. Within this context, the authors describe the organisation of VPH services for health and agriculture, which have a part to play in intersectorial collaboration and in social participation. Finally, an account is given of progress in the control of rabies, brucellosis and tuberculosis, and the organisation of integrated programmes for food protection and also the control and eradication of foot and mouth disease. Technical cooperation with the Pan American Health Organisation (PAHO) assists these activities and intersectorial coordination, principally between health and agriculture.

Abstract: A 69-year-old man living in Florence reported fever and acute lumbar pain one month after transurethral resection of a superficial transitional cell carcinoma of the bladder. The radionuclide bone scan suggested metastatic lesions of the L3-L4 vertebrae. However cobalt treatment was ineffective. A bone biopsy of L4 showed an inflammatory pattern and antibiotic therapy was started which did not produce any clinical improvement. Six months after the onset of the back pain brucellar spondylitis was serologically diagnosed and treatment with doxycycline and streptomycin produced a significant clinical and radiological improvement. After 2 months the patient's wife presented with fever and lumbar pain, and brucellar spondylitis was diagnosed as well. An extensive epidemiological examination revealed that 8 months earlier the family had eaten unpasteurized goat cheese and serological examination of the entire family showed that 3 out of 4 members had significant titres of brucellar antibodies. Finally it was discovered that 4 months after consuming the cheese the third infected subject experienced an episode of epidydimoorchitis for which no diagnosis and effective treatment was found. This family cluster of brucellar infection indicates that a high degree of suspicion in the diagnosis of brucellosis is necessary even in non-endemic areas, to reduce the delay in the diagnosis and treatment of the disease and to prevent the occurrence of complications that may prove difficult to treat.

Abstract: Brucellosis is an important zoonosis, and serological surveillance is essential to its control. However, cross-reactions of attenuated live cells of Brucella abortus strain S-19 and B. melitensis strain Rev-1 with Yersinia enterocolitica O9 or vaccinated animal sera interfere with accurate serological diagnosis by the Rose Bengal test (RBT). Therefore, we used ELISA with sarcosine extracts from the virulent B. abortus strain 544 to eliminate false-positives among RBT positive-sera. A total of 697 serum samples were collected in Mongolia from humans and animals in 23 nomadic herds. The herds were classified into three groups as brucellosis-endemic (BE), brucellosis-suspected (BS), or Brucella-vaccinated (BV). The number of 295 animals (43.0%) was positive by RBT, but 206 (69.8%) of these were positive according to ELISA; therefore, 30.2% of the RBT-positive sera were found to be false positives. The false positive samples for RTB represent 4.1%, 27.4%, and 68.2% of the animals from the BE, BS, and BV herds, respectively. In addition, 32% of RBT-positive human sera were also false positives. Thus, our ELISA would be more specific than RTB and useful for epidemiological surveillance for brucellosis.

Abstract: All 137 brucellosis cases occurring in Tabuk Province, Saudi Arabia in 1997 were studied retrospectively. Brucella agglutination titre of > or = 1/80, or rising titre plus history of typical signs and symptoms were considered evidence of infection. The incidence rate was 34/100,000, mean age 33.8 +/- 13.9 years (range: 3-72 years) and male:female ratio 1.8:1. There were 63.5% of cases rurally resident, 58.4% kept animals at home or elsewhere, 27.0% worked with animals and/or on farms, and 88.3% reported a history of raw milk ingestion. The most common infecting agents were Brucella melitensis, B. abortus and B. suis. Splenomegaly and hepatomegaly were detected in 25.5% and 22.6% of cases respectively.

Abstract: Protein assemblies with a high degree of repetitiveness and organization are known to induce strong immune responses. For that reason they have been postulated for the design of subunit vaccines by means of protein engineering. The enzyme lumazine synthase from Brucella spp. (BLS) is highly immunogenic, presumably owing to its homodecameric arrangement and remarkable thermodynamic stability. Structural analysis has shown that it is possible to insert foreign peptides at the ten amino terminus of BLS without disrupting its general folding. These peptides would be displayed to the immune system in a highly symmetric three-dimensional array. In the present work, BLS has been used as a protein carrier of foreign peptides. We have established a modular system to produce chimeric proteins decorated with ten copies of a desired peptide as long as 27 residues and have shown that their folding and stability is similar to that of the wild-type protein. The knowledge about the mechanisms of dissociation and unfolding of BLS allowed the engineering of polyvalent chimeras displaying different predefined peptides on the same molecular scaffold. Moreover, the reassembly of mixtures of chimeras at different steps of the unfolding process was used to control the stoichiometry and spatial arrangement for the simultaneous display of different peptides on BLS. This strategy would be useful for vaccine development and other biomedical applications.

Abstract: To survey the antibiotic susceptibility patterns of some commercially available Lactobacillus, we collected four commercial products that contain active Lactobacillus. We incubated individual product and identified these colonies by the methods of API50 CH test kit and RAPID ID 32A kit. Strains of Streptococcus thermophilus, Bifidobacterium infantis, Lactobacillus acidophilus and Lactobacillus casei were collected. By agar dilution method, each identified strain was inoculated to Brucella blood agar-MIC plates. Each plate contained one of the following antibiotics with different concentrations: amoxicillin, cephalothin, gentamicin, vancomycin, erythromycin, rifampin, tetracyclin and penicillin G, clindamycin, chloramphenicol, cefmetazole, metronidazole, ampicillin/sulbactum, cefoxtin, etc. After incubation, the growth condition of each Brucella blood agar-MIC plate was observed and the breakpoint of each antibiotic to different Lactobacillus products determined. The MICs of amoxicillin, ampicillin/sulbactum and penicillin-G to all identified strains were < or =2 microg/ml and those of vancomycin, clindamycin, erythromycin, metronidazole, cefmetazole and cefoxtin for L. casei were >32 microg/ml. L. casei was more resistant to all the testing antibiotics than the other strains. According to the MICs of the above antibiotics, proper active lactobacillus products could be chosen to prevent antibiotic-associated diarrhea in the pediatric field.

Abstract: The aim of this study was to investigate seroprevalences to zoonotic pathogens in hunters, to compare the results with other predisposed occupational groups already investigated and to propose preventive measures. Blood samples were taken from 146 male and 3 female hunters from the provinces of Styria and Burgenland in the south-east of Austria and anamnestic data were obtained using a questionnaire. The serological investigations included the following bacterial, viral and parasitic zoonotic agents or zoonoses, respectively (antibody prevalence rates in brackets): borreliosis (IgG 42%, IgM 7%), brucellosis (1%), chlamydiosis (3%), ehrlichiosis (IgG 15%, IgM 3%), leptospirosis (10%), tularaemia (3%), Q fever (0%), encephalomyocarditis virus (EMCV, 15%), Puumala-Hantavirus (10%), Newcastle Disease virus (4%), Echinococcus multilocularis/E. granulosus (5%/11%), toxocariasis (17%). Particularly striking in comparison with the control group and the veterinarians, farmers and slaughterhouse workers examined in earlier projects were the high seroprevalences to Borrelia burgdorferi sensu lato, Ehrlichia spp., Leptospira interrogans, E. granulosus and E. multilocularis, encephalomyocarditis, Puumala-Hantavirus and Newcastle Disease virus as well as to Brucella abortus and Francisella tularensis. The present study indicates that hunters are especially exposed to zoonotic pathogens.

Abstract: The diagnosis of bovine brucellosis using PCR in blood and milk samples from two dairy herds were compared to in vitro isolation, complement fixation test (CF), competitive ELISA (C-ELISA) in serum, and indirect ELISA (I-ELISA) in milk. Samples were obtained from 99 cows vaccinated with Brucella abortus strain 19, from a naturally infected herd (A), whose cows were also vaccinated with B. abortus strain RB51 as adults, and 100 from brucellosis free herd (B). In herd A, PCR identified 14 B. abortus infected cows: nine infected with wild type, and five with wild type and RB51, B. abortus S 19 was not identified. B. abortus biotype 1 was isolated from one cow. All cows infected with a wild strain of B. abortus were positive in serologic tests. Brucella was not found in herd B using PCR. Serological test showed 100% sensitivity related to PCR. The specificity for CF, C-ELISA and I-ELISA was 100%, 99% and 95% respectively. PCR could be useful to identify Brucella biotypes and to complement serologic tests.

Abstract: Brucellosis is being reported with increasing frequency in the Islamic Republic of Iran. Serum antibodies in high-risk and general populations help to define cut-off levels and can be used as a simple and rapid diagnostic tests in infected areas. We performed the rose Bengal test (RBT), serum agglutination test (SAT) and 2-mercaptoethanol (2ME) titre determination on 415 healthy individuals including butchers, slaughterers and others. Positive results were found by RBT, SAT titre (1:80) and 2ME titre > or = 1:20 in slaughterers (10%, 20% and 6% respectively), butchers (6%, 4% and 1% respectively) and the general population (1%, 2% and < 1% respectively). A single SAT titre > or = 1:80 in the presence of 2ME titre > or = 1:20 can be diagnostic in this region.

Abstract: It is a dogma, that RB51 vaccination does not induce antibodies that interfere with Brucellosis diagnosis, therefore any animal positive to serological test is considered as an infected animal. To determine protection against Brucellosis virulent field strain, 35 pregnant cows from a free-Brucellosis herd, previously vaccinated as calves with 1 x 10(10) CFU of RB51, were revaccinated with RB51 reduced dose, and then introduced into a herd with an active outbreak. Seventeen cows resulted positive in card test after revaccination. All 35 pregnant revaccinated cows had normal parturition; nevertheless, RB51 vaccine strain was isolated from milk and vaginal exudates from two cows after delivery at day 120 post-revaccination. At 150 days post-revaccination, two cows were positives to card and rivanol test and the field virulent strain was isolated. Revaccination with a reduced dose of RB51 in endemic zones did not cause abortion and protected 94% of animals against field infection, but caused an atypical response to conventional serological tests.

Abstract: OBJECTIVE: To identify the risk factors for brucellosis in Palestine, to estimate their association with the disease and to recommend appropriate prevention measures. METHODS: An unmatched case-control study was conducted in the West Bank of Palestine. A total of 450 subjects (150 cases and 300 controls) chosen from all districts were investigated. Data were collected during the year 2000. Subjects were interviewed using a standard questionnaire acquiring demographic and risk factor information. Laboratory results were also recorded on the questionnaire. Data were analyzed calculating the odds ratio and the confidence intervals for the studied variables. A logistic regression model was used to explore the association between disease status and the studied variables. RESULTS: Several variables were significantly associated with brucellosis involving both direct and indirect transmission such as consumption of unpasteurized milk and dairy products, herding, lambing and others. CONCLUSION: Most of the risk factors for being infected with brucellosis are modifiable. Efforts should be directed to the prevention of this major public health problem in Palestine utilizing the information obtained in this study.

Abstract: OBJECTIVES: The aim of our study was to analyse the antibiotic susceptibility of various strains of Bifidobacterium spp. to a wide range of antimicrobial agents. METHODS: Fifty strains belonging to eight species of bifidobacteria, isolated from humans, animals or probiotic products, were tested for susceptibility to 30 antibiotics by disc diffusion on Brucella agar supplemented with 5% laked sheep blood and vitamin K1 (1 mg/L). MICs of nine anti-anaerobe agents, including three new molecules (telithromycin, linezolid and gatifloxacin), were determined using the reference agar-dilution method. RESULTS: All strains of bifidobacteria, whatever the species, were sensitive to penicillins: penicillin G, amoxicillin (MIC(50) 0.06 mg/L), piperacillin, ticarcillin, imipenem and usually anti-Gram-positive antibiotics (macrolides, clindamycin, pristinamycin, vancomycin and teicoplanin). Susceptibility to cefalothin and cefotetan was variable. Most isolates (70%) were resistant to fusidic acid. As expected, high resistance rates were observed for aminoglycosides. Metronidazole, an agent known for its anti-anaerobe activity, was ineffective against 38% of the strains. The newly commercialized molecules, telithromycin, linezolid and gatifloxacin, were active with MIC(50)S of 1 mg/L. The only variation in susceptibility observed among the different species concerned Bifidobacterium breve, which appeared to be generally more resistant. Potentially acquired resistance was only observed against tetracycline and minocycline, in 14% of the strains. CONCLUSIONS: With regard to a general concern about the safety of probiotics, such as potential transferability of resistance determinants, bifidobacteria, with their low natural and acquired resistance to 30 antibiotics, appear risk-free.

Abstract: The epidemiological features and clinical manifestations of adult cases of brucellosis admitted to the Department of Infectious Diseases, Babol Medical University, Iran from 1997 to 2002 were investigated. Of 469 cases, 267 (56.9%) were males. The mean age of cases was 36.9 +/- 15 years. Most (60.8%) were from rural areas. Two thirds of cases (306, 66.3%) presented during spring or summer. Fresh cheese (22.4%), animal husbandry (11.3%), laboratory worker (8.1%) and veterinary profession (1.5%) were the main risk factors. Forty-five families (9.6%) had two cases. Sweating, fever, and arthralgia were the most frequent clinical symptoms. Complications were documented in 105 males (39.5%) and 41 females (20.3%, P=0.0001). Peripheral arthritis was seen in 24 (9%) males and 19 (9.4%) females, with knees and hips being the most common sites of infection. Sacroiliitis and spondylitis were seen in 28 (6%) and 32 (6.8%) cases respectively with spondylitis more common in males (P=0.023). Epididymo-orchitis was seen in 29 (10.9%) males. There were three cases each of endocarditis (0.6%) and neurological complications (0.6%). Most patients with brucellosis did not have any of the known risk factors for brucellosis. Thus consumption of unsafe dairy products could be the main route of infection. The disease manifested with a diversity of clinical manifestations and complications. Complications were more frequent in males than females.

Abstract: A 15-month-old boy was admitted with fever and a swollen knee. His mother had been treated for brucellosis 11 months ago. At that time he had been asymptomatic and had both negative blood culture and serum agglutination tests and breastfeeding had been stopped. The infant had been healthy since then. On admission, blood and joint fluid were obtained for culture and he was commenced on cefuroxim. Gram-negative coccobacillary organisms were seen in the joint fluid. Both cultures remained sterile. He had a positive serum agglutination test with a titer of 1/640. Cefuroxim was then stopped and the child was commenced on a specific course of treatment: gentamicin, trimethoprim-sulfamethoxazole and rifampicin. The infant recovered with this treatment. We conclude that this was brucellar arthritis of the knee and was probably acquired by breastmilk after an exceptionally long incubation period.

Abstract: Milk culture data and serologic test results were evaluated after adult vaccination with Brucella abortus strain 19 in cattle of 3 large California dairy herds infected with brucellosis. Strain-19 organisms were isolated by culture of milk from 1.9% of the vaccinated cows. Isolation of field strain of B abortus varied directly with magnitude of complement-fixation (CF) and rivanol titers. At time of milk culture, 74% of cows from which field strain was isolated had CF titer greater than or equal to 160, compared with 58% of cows from which strain 19 was isolated. Cows with CF titer greater than or equal to 160 at 2 months or greater than or equal to 80 to 4 months after adult vaccination were more likely to be correctly classified as reactors (on the basis of subsequent milk culture results and/or persistently high serologic titer) than were cows with lower CF titer at these times. Cows from which B abortus strain 19 was isolated from milk were more likely to maintain persistent serologic titer than were cows from which neither strain of B abortus was isolated.

Abstract: Brucella melitensis is the cause of brucellosis in sheep and goats, which often results in abortion. Few cases of B. melitensis infection in goats have occurred in the United States over the last 25 years. However, vigilance must be maintained, as it is for the bovine milk industry, to ensure that brucellosis is not introduced into the U.S. goat population. The objective of this study was to develop a sensitive and specific indirect enzyme-linked immunosorbent assay (iELISA) for the detection of B. melitensis-specific antibodies in goat milk. Brucella salt-extractable protein extract was employed as an antigen, and a horseradish peroxidase-labeled polyclonal anti-goat antibody was used as an anti-species conjugate. Thirteen of 13 (100%) individual infected goat milk samples tested positive and 134 of 134 (100%) uninfected bulk milk samples tested negative by the developed iELISA. Three positive milk samples with high, medium, and low absorbance values were used to simulate one positive animal in an otherwise negative herd. By this estimation, one high-titer animal could be detected in a herd of >1,600 animals. Detection estimates for medium- and low-titer animals were one positive animal per herd of <200 and 50 animals, respectively. Based on this estimation, it is recommended that herds be sampled in groups of 50 animals or less for bulk milk testing. The iELISA developed for this study was found to be sensitive and specific and shows potential for use as a bulk milk test for the detection of B. melitensis-specific antibodies in goat milk.

Lopetegui P. Bovine brucellosis control and eradication programme in Chile: vaccine use as a tool within the programme. Developments in biologicals. 2004; 119; 473-9. [PubMed: 15742662].

Abstract: In 1975 Chile began the bovine brucellosis control programme in the south central area of the country, where the 92% of the bovine population is located. In 1992 the programme was evaluated and the prevalence showed almost no change for the last nine years, Therefore, a comprehensive Brucellosis eradication programme began. Based on the information gathered by a surveillance system on cattle market and dairy herds from 1999 to 2003 the advances in the process are shown. By applying this programme it has been possible to eradicate brucellosis from the XII Region, and in 2003 was declared brucellosis free with vaccination. In Region XI the project is in its final year and during the last semester no reactor animals have been found. The central south regions began their eradication in 1996. This is the area of the country where the largest bovine population is concentrated, and also the highest brucellosis prevalence. Now the situation has been changing and the trend shows fewer reactors in auction markets and milk plants. The effect of the vaccine (Strain 19 at the beginning and Strain RB 51 from 1997) has been important in impeding the infection of negative herds that are in infected areas. It is also used with success in infected herds, applying "whole herd vaccination" to create a quick herd immunity. This has been important since in Chile there is no paid compensation for reactor animals and the infected animals remain for a variable period in the herd before being destroyed.

Abstract: We determined the prevalence of tuberculosis and brucellosis reactors in the dairy herds in the Mbarara district of Uganda in 2002. This is one of the most important dairy-production areas of the country and includes both pastoral and agro-pastoral zones. A total of 340 (of 11,995) randomly selected herds were tested for tuberculosis, using the intradermal tuberculosis-skin test and 315 (of 10,562) herds tested for brucellosis using the serum Rose Bengal test. The herd prevalence for tuberculosis reactors was 74.1% (95% confidence intervals 69, 78), the individual-animal prevalence was of 6.0% (5.6, 6.5) and within-herd range was 1-50% (up to 100% if suspicious reactors were included). The herd prevalence for brucellosis was 55.6% (50, 61.2) individual-animal prevalence 15.8% (14.8, 16.7) and within-herd range 1-90%. The reactor prevalence increased with the age of the animals for both tuberculosis and brucellosis. Tuberculosis reactor prevalences were higher in animals from the agro-pastoral zone. However, the individual-animal and herd prevalences of brucellosis seroprevalences were higher in the pastoral zone.

Abstract: Sinorhizobium meliloti, a legume symbiont and Brucella abortus, a phylogenetically related mammalian pathogen, both require their BacA proteins to establish chronic intracellular infections in their respective hosts. The lipid A molecules of S. meliloti and B. abortus are unusually modified with a very-long-chain fatty acid (VLCFA; C > or = 28) and we discovered that BacA is involved in this unusual modification. This observation raised the possibility that the unusual lipid A modification could be crucial for the chronic infection of both S. meliloti and B. abortus. We investigated this by constructing and characterizing S. meliloti mutants in the lpxXL and acpXL genes, which encode an acyl transferase and acyl carrier protein directly involved in the biosynthesis of VLCFA-modified lipid A. Our analysis revealed that the unusually modified lipid A is important, but not crucial, for S. meliloti chronic infection and that BacA must have an additional function, which in combination with its observed effect on the lipid A in the free-living form of S. meliloti, is essential for the chronic infection. Additionally, we discovered that in the absence of VLCFAs, S. meliloti produces novel pentaacylated lipid A species, modified with unhydroxylated fatty acids, which are important for stress resistance.

Abstract: Although aggressive public health measures have greatly reduced the number of brucellosis cases in the United States, there is a resurgence of interest in this worldwide zoonosis because of its potential as a bioweapon and its 8-fold higher incidence in California, Texas, and the other borderlands between the United States and Mexico compared with the national rate. Accordingly, we reviewed the clinical records of 28 patients diagnosed at a university hospital in San Diego, CA, between 1979 and 2002 to look for new epidemiologic trends and to test the hypothesis that there are species-specific differences in clinical presentations. In contrast to the latest California-wide study completed in 1992, Brucella abortus infections were more common (73%) than Brucella melitensis after 1992, and women were more commonly infected (77% compared with 39%) than men. Major risk factors remained Hispanic ethnicity, travel to Mexico, and ingestion of nonpasteurized dairy products. Analysis of diagnostic procedures suggested that the traditional practice of prolonged incubation of blood cultures increased their sensitivity for Brucella, even in automated radiometric systems. Direct comparison of the clinical manifestations of infections with B. abortus and B. melitensis strongly supported differences in acute presentations. B. melitensis presented more acutely as fevers of unknown origin with statistically significant higher rates of abdominal tenderness, hepatomegaly, splenomegaly, thrombocytopenia, pancytopenia, and hepatic dysfunction. These results suggest that the epidemiology of brucellosis in California may be evolving, and they show, to our knowledge for the first time in a single series, that species-specific differences in presentations may account for some of the protean manifestations of brucellosis. Familiarity with manifestations of brucellosis and the optimal laboratory techniques for its diagnosis could help physicians protect the public against this reemerging, under-recognized zoonosis.

Abstract: OBJECTIVES: To evaluate the efficacy of gentamicin-loaded poly (lactide-co-glycolide) 50:50H (PLGA 50:50H) microspheres for the treatment of mice experimentally infected with Brucella abortus 2308. METHODS: The microspheres were dispersed in either 2% (w/v) poloxamer 188 saline solution, or deionized water with the help of a cell homogenizer to break up particle aggregates, and were administered intravenously or intraperitoneally to B. abortus-infected mice 7 days post-infection. RESULTS: Neither a single intravenous or intraperitoneal dose of 67 microg of gentamicin per mouse, nor three intraperitoneal doses of 100 microg of gentamicin per mouse, reduced the Brucella infection in the spleen compared with untreated mice 1 and 3 weeks post-treatment. Histological examination revealed granulation and tissue reaction in the periphery of spleen and liver of animals given three doses of the gentamicin-loaded microspheres. CONCLUSIONS: The lack of therapeutic activity of the gentamicin-loaded microspheres might be related to inappropriate microsphere size and aggregation, resulting also in a poor distribution of the microspheres in the spleen. The results might provide an example of practical problems related to particle size and aggregation for in vivo therapy with PLGA microspheres.

Abstract: Milk and blood samples from 704 lactating ewes were examined for the diagnosis of Brucella melitensis infection by milk-ELISA, serum-ELISA, RBPT, SAT and culture of milk. Of these ewes, 209 were from brucellosis free sheep flock, 443 from brucellosis infected sheep flock and 52 were from private sheep flocks of which status for brucellosis was not known. All the 209 ewes belonging to uninfected sheep flock were found negative in all the tests and of the remaining 495 ewes 105 were positive in serum-ELISA, 103 in milk-ELISA, 92 in RBPT, 85 in SAT, and B. melitensis biovar-1 was isolated from the milk of 29 ewes. Of the 105 serum-ELISA positive ewes, 99 were positive and 6 were negative in milk-ELISA, whereas of the 103 milk-ELISA positive ewes, 4 were negative in serum-ELISA. All together, 99 ewes were positive and 386 were negative in both the assays while 10 ewes yielded variable results. The specificity of milk-ELISA in brucellosis free flock was 100% and sensitivity and positive predictive value were 96.11% and 94.28%, respectively, in infected flocks. The Brucella antibody levels in milk and serum samples as determined by milk-ELISA and serum-ELISA were correlated significantly. The milk-ELISA for brucellosis appears to be an attractive alternative of serum-ELISA particularly in the lactating ewes.

Abstract: Two hundred and fifty human serum samples were collected (122 from general population and the rest from people associated with animals) from central Kerala and tested for Brucella agglutinins using various standard tests. The overall seropositivity for brucellosis using all the tests was 1.6%. Among the general population a prevalence of 2.45% was observed and among the veterinary students 1.14%.

Abstract: BACKGROUND: Brucellosis is a world-wild zoonosis of bacterial origin. In Italy, where reporting of the disease is mandatory, the incidence is 2.1 cases every 100,000 inhabitants, but tends to be higher in southern regions. However, the incidence decreased from 1999 to 2002. CASES: Five cases of occupational brucellosis, four slaughter-house operators and one veterinary surgeon, were reported in a slaughter-house in south-eastern Sicily employing 15 people. RESULTS: A study of the slaughtering process for sheep and goats led to identification of 23 different operations, some of which involved greater probability of infection: animal consignment; unloading and leading animals from the stable to the trap; strangling and bleeding; skinning and removal of mammary glands; removal and processing of abdominal viscera; post-mortem examination; mobile equipment and facility maintenance, and washing/disinfection. All affected workers had participated in one or more of these operations. CONCLUSION: Eradication of brucellosis in stock-farms should lead to the disappearance of the human disease, since the two are closely related. Despite legislation aimed at eradicating brucellosis in cattle, which has been in force since 1992, the prevalence of the human disease has risen in Italy, especially in Sicily. Within the framework of prevention, occupational physicians should exercise specific surveillance of brucellosis risk by carefully examining the plasma antibody content of workers involved in the slaughtering process. Occupational health physicians should also provide ad hoc information regarding the infectious agent and modes of transmission during slaughtering operations as well as specific training in the use of protective equipment, which is the only way of providing protecting from contagion by infected animals.

De Massis F, Di Girolamo A, Petrini A, Pizzigallo E, Giovannini A. Correlation between animal and human brucellosis in Italy during the period 1997-2002. Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases. 2005 Aug; 11(8); 632-6. [PubMed: 16008615].

Abstract: The aim of this study was to test the hypothesis that brucellosis in Italy is a food-borne, rather than an occupational disease. This hypothesis was tested using data for both human and animal populations from the period 1997-2002. The correlation between the distribution of the disease in the human, sheep and goat populations was analysed, as were the risk factors for the disease, with respect to gender, age, occupation and residence of the individuals involved. Notifications of human brucellosis, which are mandatory in Italy, reach a peak between April and June. However, considering the standard incubation period of 2-4 weeks, and the fact that lamb slaughter is traditionally at a peak during the Easter period, it might be expected that occupational exposure would result in a peak of human cases between March and May. The observed peak between April and June could be related to the production and consumption of fresh cheese, starting just after lamb slaughter. The age of patients showed a fairly uniform distribution, and analysis of incidence rates of human brucellosis between 1997 and 2002 showed that the incidence rates were consistent with an occupational exposure risk of about 25%.

Abstract: Genes thuA and thuB in Sinorhizobium meliloti Rm1021 code for a major pathway for trehalose catabolism and are induced by trehalose but not by related structurally similar disaccharides like sucrose or maltose. S. meliloti strains mutated in either of these two genes were severely impaired in their ability to grow on trehalose as the sole source of carbon. ThuA and ThuB show no homology to any known enzymes in trehalose utilization. ThuA has similarity to proteins of unknown function in Mesorhizobium loti, Agrobacterium tumefaciens, and Brucella melitensis, and ThuB possesses homology to dehydrogenases containing the consensus motif AGKHVXCEKP. thuAB genes are expressed in bacteria growing on the root surface and in the infection threads but not in the symbiotic zone of the nodules. Even though thuA and thuB mutants were impaired in competitive colonization of Medicago sativa roots, these strains were more competitive than the wild-type Rml021 in infecting alfalfa roots and forming nitrogen-fixing nodules. Possible reasons for their increased competitiveness are discussed.

Abstract: As part of a study to assess zoonotic milk-borne health risks, seasonal survey data and unpasteurized milk samples were collected between January 1999 and February 2000 from randomly selected informal milk market agents (220 and 236 samples in the dry and wet seasons, respectively) and from households purchasing raw milk (213 and 219 samples in the dry and wet seasons, respectively) in rural and urban locations in central Kenya and screened for antibodies to Brucella abortus (B. abortus) and presence of Escherichia coli (E. coli) O157:H7. The latter was assessed based on samples from consumer households only. Antibodies to B. abortus were screened using the indirect antibody Enzyme Linked Immunosorbent Assay (ELISA) and the Milk Ring Test (MRT). The presence of E. coli O157:H7 was assessed by culture, biochemical characterisation, serological testing for production of verocytotoxin one (VT1) and two (VT2) and polymerase chain reaction (PCR) analysis for the presence of genes encoding for the toxins. The prevalence of antibodies to B. abortus varied considerably ranging from none in milk sold in small units and originating from intensive production systems to over 10% in samples that were bulked or originating from extensive production systems. E. coli O157:H7 was isolated from two samples (0.8%), one of which produced VT1. All urban consumers (100%) and nearly all rural consumers (96%) of marketed milk boiled the milk before consumption, mainly in tea, thus greatly reducing chances of exposure to live pathogens and potential health risks.

Abstract: This study was conducted to determine the seroprevalence of human brucellosis and identify the potential risk factors in a rural area of Western Anatolia, Turkey. A simple random-sampling method was used for identifying 1,052 subjects for the study. Blood samples, collected from all the subjects, were studied following the methods of Rose Bengal slide agglutination and standard tube agglutination tests. One thousand and one samples (95.2%) were seronegative, and 51 (4.8%) were seropositive. There was a statistically significant correlation between seropositivity and age, sex, consuming fresh cheese and cream made from unboiled milk (p values 0.005, 0.019, <0.001, and <0.001 respectively). Seropositivity was not related to educational level (0.270). It is concluded that pasteurization of milk and dairy products and education regarding eating habits must be pursued for eradication of human brucellosis from rural areas. The findings of the study suggest that human brucellosis is still an important public-health problem in the western Anatolia region of Turkey, especially in rural areas.

Abstract: Brucellosis is an important zoonosis throughout the world. Infection is transmitted by the consumption of non-boiled or non-pasteurized milk and milk products such as cheese, cream and butter. The aim of this study was to investigate the presence of Brucella spp. in the fresh cheese samples which were sold at bazaars of district in Sivas center during April, May and June of the years 2003 and 2004. In 3 (7.1%) of 42 cheese samples collected in 2003 and 4 (8.5%) of 47 samples collected in 2004, Brucella spp. were found to be positive. Consequently, since the fresh cheese samples are the most important routes of Brucella transmission, the risk for public health should be well considered.

Abstract: This study compared responses of bison calves to 10(10)CFU of Brucella abortus strain RB51 (SRB51) delivered by parenteral or ballistic methods. Two types of biobullet payloads were evaluated; compacted SRB51 pellets or SRB51 encapsulated in photopolymerized poly(ethylene glycol) hydrogels. Bison were vaccinated with saline, parenteral SRB51 alone, or in combination with Spirovac, or ballistically with compressed SRB51 or hydrogel biobullets. Bison parenterally vaccinated with SRB51 had greater (P<0.05) immunologic responses when compared to control bison. Co-administration of Spirovac as an adjuvant did not influence immunologic responses. As compared to compressed SRB51 biobullets, ballistic vaccination with hydrogel biobullets increased cellular immune responses at some sampling times. Our data suggest that hydrogel formulations of SRB51 may be a superior alternative to compressed SRB51 tablets for ballistic vaccination of bison. Although preliminary, data suggests that immunologic responses of bison to SRB51 hydrogel bullets are similar to responses after parenteral vaccination with SRB51.

Abstract: Photopolymerized poly(ethylene glycol) (PEG)-crosslinked hydrogels were assessed for their ability to serve as a payload vehicle to deliver a viable bacterial vaccine (Brucella abortus strain RB51 (RB51) to bison in Yellowstone National Park) ballistically using thermoplastic degradable Biobullets. PEG modified with degradable glycolide or lactide oligomers capped with photopolymerizable methacrylate groups served to crosslink the hydrogel vaccine carrier inside commercial hydroxypropylcellulose Biobullets. Release of 1 microm diameter model fluorescent particles from hydrogels followed known degradation trends for glycolide- and lactide-modified PEG hydrogels. All particles were released from PEG-co-glycolide hydrogels after approximately 10 days and PEG-co-lactide hydrogels after approximately 45 days following gel degradation. Minimal particle release was observed from pure PEG dimethacrylate hydrogels over 40 days. P. aeruginosa (strain PAO1) and RB51 live vaccines exhibit excellent viability following exposure to photopolymerization encapsulation within these gel matrices. Hydrogels photopolymerized into the payload chamber of Biobullets exhibit similar ballistic properties to commercially available Biobullets and penetrate and remain intact when fired intramuscularly into live elk for release of their gel payload in the host.

Abstract: Brucella broth without supplementation is the recommended medium for broth microdilution susceptibility tests of Brucella abortus, B. melitensis, and B. suis. Based on an eight-laboratory collaborative study using a pH-adjusted modification of this medium, we propose MIC quality control ranges for three control strains against 10 antimicrobials that are potentially efficacious for treating infections caused by these agents of bioterrorism.

Abstract: Yersinia enterocolitica and Yersinia pseudotuberculosis have been identified as causative organisms of reactive arthritis in humans. We evaluated a Western blot assay which uses Yersinia outer membrane proteins as antigens for the detection of Yersinia antibodies as a replacement for the complement fixation (CF) assay. Clinical agreement, sensitivity, and specificity were determined by testing 19 positive and 21 negative serum samples by the CF assay, Western blot assay, and enzyme-linked immunosorbent assay (ELISA). The CF assay and ELISA were compared to the Western blot assay, which was the reference method used in this study. Sera with antibodies that could potentially cross-react with Yersinia were also tested by the Western blot assay. The agreement, sensitivity, and specificity of the CF method were 61%, 26%, and 95%, respectively; and those for the ELISA were 89%, 95%, and 82%, respectively. The prevalences of Yersinia antibodies in 50 healthy donors were 6% for immunoglobulin G (IgG), 2% for IgA, and 2% for IgM. Sera positive for Bartonella henselae, Brucella, Chlamydia pneumoniae, and Rickettsia rickettsii antibodies showed cross-reactivity by the Western blot assay. The highest cross-reactivity was observed with Borrelia burgdorferi; 5 of 11 (45%) specimens were cross-reactive by the IgM-specific assay. Overall, the Western blot assay performs acceptably and is more sensitive than the CF assay, warranting replacement of the CF assay in the laboratory. Due to the evidence of cross-reactivity, particularly with B. burgdorferi, which can cause an oligoarthritis similar to reactive arthritis, the diagnosis of reactive arthritis should be based on clinical findings and complete serologic analysis of the potential causative infectious pathogens.

Abstract: The authors studied the persistence of infection in 46 ewes experimentally infected with Brucella melitensis biovar 3 and monitored through three subsequent reproductive cycles. The entire experimental period lasted for 151 weeks. Infection of ewes and elimination of Brucella in milk, or its presence in vaginal discharges, persisted throughout the duration of the trial, as demonstrated by recurrent elimination of Brucella in milk and vaginal discharges. Brucella melitensis was recovered from the tissues of one ewe killed at the end of the trial. The strain was recovered from vaginal swabs and milk following parturition in the third reproductive cycle from an ewe that had aborted in the first cycle but was not pregnant in the second cycle. From a public health point of view, the periodical recovery of Brucella from the milk during the entire trial period illustrated that brucellosis in sheep remains a continuous occupational risk and a significant public health problem for consumers of fresh milk and milk products. That risk may persist for at least 3 years following the initial infection of the flock. Lamb antibody titres became negative in all lambs within 5 months after birth. This suggested that serological tests on lambs may have no practical diagnostic significance if performed during the first 5 months of life. Nevertheless, the birth of three infected lambs suggested that the phenomenon of latent carrier state may represent another way for B. melitensis to persist in a flock.

Abstract: The latex agglutination immunoassay technique uses polymer colloids as carriers for antibodies or antigens to enhance the immunological reaction. In this work, the interaction of a lipopolysaccharide (LPS) of Brucella Melitensis with two conventional latexes has been studied. Some experiments on the physical adsorption of the LPS onto these polystyrene beads have been performed and several complexes with different coverage degrees were obtained by modifying the incubation conditions. Regarding the application in the development of diagnostic test systems, it is advisable to study the latex-LPS complexes from an electrokinetic and colloidal stability point of view. The complexes were electrokinetically characterized by measuring the electrophoretic mobility under different redispersion conditions. The colloidal stability was determined by simple turbidity measurements. Experimental and theoretical data have been employed to study the molecular disposition of the LPS in the latex particle surface to compare with the outer membrane of bacterial cells. Latex complexes covered by different LPS amounts showed high colloidal stability and adequate immunoreactivity that remains for a long time period.

Abstract: Due to the important drawbacks of the Brucella melitensis Rev 1 vaccine, a safer vaccine based on an outer membrane complex from Brucella ovis encapsulated in poly-epsilon-caprolactone (PEC) microparticles (MP) was developed and tested in rams. Homogeneous batches of microparticles were prepared by a new double emulsion solvent evaporation method called "Total Recirculation One-Machine System" (TROMS). Such microparticles presented a mean diameter of 2 microm and displayed an antigen loading of about 13 microg HS per mg of microparticles. Subcutaneous vaccination of rams with 800 microg HS (hot saline antigenic extract of B. ovis) in PEC microparticles induced an adequate serological response against B. ovis antigens and conferred similar protection against challenge with B. ovis to that induced by the living attenuated B. melitensis Rev 1 reference vaccine. By contrast, lower doses (80 microg) of HS-PEC evoked reduced serological responses against B. ovis antigens and did not induce significant protection. The revaccination with 800 microg of HS-PEC increased the intensity and duration of the serological response against B. ovis antigens but did not improve the protection conferred by the single vaccination. Sample sera taken from any of the animals immunized with Rev 1 were seropositive in both Rose Bengal and the Complement Fixation tests (RBT, CFT) used for the diagnosis of smooth Brucella infections. By contrast, no positive reactors in both tests were recorded in the animals vaccinated with HS-PEC, being this a target objective of this study. HS-PEC microparticles can be used as a safe vaccine against brucellosis in rams, but further studies using higher doses of antigens are necessary to exploit their full potential for the prophylaxis of brucellosis in sheep.

Abstract: Eleven cases of human brucellosis occurred among livestock workers and a veterinarian who lived and worked in a rural area around Jeongeup City, Jeollabuk-Do, Korea from February 2003 to August 2003. Eight of the patients had taken care of Korean native cattle that were infected with bovine brucellosis and had already been slaughtered. Two of the patients had taken care of dairy cattle, and one case was a veterinarian who acquired the disease through an accidental contact with infected cattle while assisting in calf delivery. Eleven cases were identified by serologic work ups and four cases were identified via positive blood cultures. This study shows that the Republic of Korea is no longer free of human brucellosis, Brucella abortus biotype 1. We reviewed the patients' characteristics and serologic data during the one-year follow up period, and we also discuss on the efficacy and side effects of the rifampin and doxycyline regimen used for the treatment of human brucellosis.

Abstract: Brucellosis is a significant health problem in countries where control of zoonoses is inadequate. During 1993-98, we analysed sera and cultures from 792 suspected brucellosis patients who presented with histories of fever, chills, night sweating, weakness, malaise and headache to the referral hospital in Yazd. Cases were investigated by tube agglutination test (TAT) and 2-mercaptoethanol test (2-MET) and a questionnaire was completed for each.TAT titre was > or = 1:1 60 for 745 patients (94.1%) and 2-MET was positive for 42 (5.3%). Of 745 confirmed cases, 460 were from 1996-1997. Prevalence was highest in summer (39.5%) and more common males than among females. Prevalence was highest among those aged 10-19 years (27.7%). Most patients had a history of infected cheese, milk and milk product consumption (98%).

Abstract: The enzyme lumazine synthase from Brucella spp. (BLS) is a highly immunogenic protein that folds as a stable dimer of pentamers. It is possible to insert foreign peptides and proteins at the 10 N terminus of BLS without disrupting its general folding, and these chimeras are very efficient to elicit systemic and oral immunity without adjuvants. In this study, we show that BLS stimulates bone marrow dendritic cells from mice in vitro to up-regulate the levels of costimulatory molecules (CD40, CD80, and CD86) and major histocompatibility class II Ag. Furthermore, the mRNA levels of several chemokines are increased, and proinflammatory cytokine secretion is induced upon exposure to BLS. In vivo, BLS increases the number of dendritic cells and their expression of CD62L in the draining lymph node. All of the observed effects are dependent on TLR4, and clearly independent of LPS contamination. The described characteristics of BLS make this protein an excellent candidate for vaccine development.

Abstract: The aim of this study was to evaluate the effect of the excipients used to facilitate the encapsulation of high hydrophobic antigenic complex extracted from Brucella ovis (HS) on the physico-chemical properties of the resulting microparticles. Poly(epsilon-caprolactone) (PEC) microparticles containing HS were prepared by the solvent extraction/evaporation method using total recirculation one-machine system (TROMS). Different excipients, beta-cyclodextrin (beta-CD), Pluronic F68, Tween 20 or Tween 80, were used in order to facilitate the encapsulation and conserve the bioactivity of the encapsulated antigenic complex. HS was efficiently loaded in all the different PEC-microparticle formulations, although the combined use of beta-cyclodextrin and Pluronic F68 permitted an increase in the amount of antigenic extract in the core of the resulting microparticles without loss of its antigenic properties. Finally, the protective ability of this F68-CD-MP formulation was evaluated against an experimental challenge with the virulent Brucella melitensis H38 strain in BALB/c mice. This innocuous subcellular vaccine induced a similar protection to that of the live attenuated Rev 1 vaccine; these are promising results that would merit further investigation in target animals.

Abstract: This study pretends to determine baseline data on the health and mortality of a colony of captive collared peccaries in the Eastern Amazon (Belém, State of Pará, Brazil) during a 65-months survey. Thirty-nine out of 166 animals (23.5%) died and were examined post-mortem. Monthly mortality averaged 1.2%. The highest mortality rate was observed in newborns (74.4%). Abandonment by the mother and aggression were responsible for 24.1% and 13.8% of the total newborn deaths, respectively. Most frequent causes of non-neonatal death were food poisoning (50.0%) due to an episode of accidental bitter cassava leaves ingestion and traumatism due to aggressions between animals (10.0%). Results from serology for different infectious diseases showed that 4.9% (2/41) collared peccaries had antibodies against Brucella spp. and 9.8% (4/41) animals had antibodies to two different Leptospira spp. serovars, butembo and autumnalis. This is the first survey of morbidity and mortality in captive collared peccaries in the Amazon region.

Abstract: Unlike Bacillus subtilis and Escherichia coli, the gram-positive lactic acid bacterium Lactococcus lactis does not possess the SecDF protein, a component of the secretion (Sec) machinery involved in late secretion stages and required for the high-capacity protein secretion in B. subtilis. In this study, we complemented the L. lactis Sec machinery with SecDF from B. subtilis and evaluated the effect on the secretion of two forms of staphylococcal nuclease, NucB and NucT, which are efficiently and poorly secreted, respectively. The B. subtilis SecDF-encoding gene was tested in L. lactis at different levels. Increased quantities of the precursor and mature forms were observed only at low levels of SecDF and at high NucT production levels. This SecDF secretion enhancement was observed at the optimal growth temperature (30 degrees C) and was even greater at 15 degrees C. Furthermore, the introduction of B. subtilis SecDF into L. lactis was shown to have a positive effect on a secreted form of Brucella abortus L7/L12 antigen.

Abstract: Brucellosis is a highly infectious disease which is diagnosed using serological and microbiological methods. The objective of this study was to assess the viability of using conventional and real-time PCR assays as potential diagnostic tools for the detection of Brucella abortus in naturally infected cows. PCR assays that amplify various regions of the Brucella genome, IS711 genetic element, 31kDa outer membrane protein and 16S rRNA, were optimised using nine known Brucella strains. Real-time PCR was used to examine the detection efficiency of the IS711 assay which was estimated at 10 gene copies. Milk, blood and lymph tissue samples were collected from naturally infected animals. B. abortus was not detected in blood samples collected from naturally infected cows by conventional or real-time PCR, but was detected in a proportion of the culture-positive milk (44%) and lymph tissue (66% - retropharyngeal, 75% - supramammary) samples by the same methods. There was no difference between PCR and bacteriological detection methods. It is unlikely that conventional or real-time PCR will supersede current diagnostic methods for detection of B. abortus in clinical samples.

Abstract: This paper reports on a sporadic, naturally acquired infection of sheep with Brucella abortus on a privately owned farm in Toro near Bauchi, Nigeria. The abortions, which occurred in a flock of 28 Yankassa sheep, involved five ewes at the third month of gestation. Serum and milk samples from the flock were examined for Brucella antibodies by the Rose Bengal plate test, serum agglutination test (SAT) and milk ring test (MRT). The proportion shown as positive by SAT was 14.3%. All the five milk samples examined by MRT were positive. A total of seven isolates of Brucella were obtained from three milk samples and four vaginal swabs collected from aborting ewes. All isolates were identified and biotyped as B. abortus biovar 1. This biovar was also isolated from cattle maintained on the farm in association with the sheep. The infection was attributed to the animal husbandry practices employed on the farm.

Abstract: This paper describes an indirect enzyme-linked immunosorbent assay (I-ELISA) and a fluorescence polarisation assay (FPA), each capable of detecting antibody in several species of hosts to smooth and rough members of the genus Brucella. The I-ELISA uses a mixture of smooth lipopolysaccharide (SLPS) and rough lipopolysaccharide (RLPS) as the antigen, and a recombinant protein A/G conjugated with horseradish peroxidase as the detection reagent. When using individually determined cutoff values, the SLPS/RLPS combined-antigen I-ELISA detected antibody in slightly more animals exposed to SLPS or to RLPS than did I-ELISA procedures using each individual antigen separately. Similarly, the assay using combined antigens detected antibody in slightly fewer animals not exposed to Brucella sp. When a universal cutoff of 10% positivity was used (relative to strongly positive control sera of each species), the overall performance index (percentage sensitivity plus percentage specificity) value decreased by 1.0 (from 199.4 to 198.4). In the FPA, it was not possible to use a universal cutoff without significant loss of performance. The overall sensitivity value for the FPA using the combined FPA antigen was 1.0% lower than using the O-polysaccharide (OPS) from SLPS and 9.1% higher than using the core antigen (CORE) from RLPS. When the combined antigen was used, the FPA specificity was slightly higher (1.2%) than from only the OPS, and considerably higher (12.6%) than the CORE. Overall, both the I-ELISA and the FPA with combined antigens were suitable as screening tests for all species of Brucella in the animal species tested.

Abstract: The diagnostic techniques most widely used for detecting brucellosis caused by Brucella ovis are serological tests such as complement fixation (CFT), agar gel immunodiffusion (AGID), and ELISAs. However, to our knowledge, milk tests, with the advantage that samples may be taken in a non invasive manner, have not been investigated as diagnostic tools. We studied 144 samples of milk and sera from lactating ewes, comparing bacteriological studies, serological and milk tests using Brucella canis and B. ovis antigens. A group of 75 ewes in an uninfected flock were serologically negative to rapid slide agglutination test (RSAT), indirect ELISA (IELISA)-B. canis, AGID and IELISA-B. ovis. The milk of these ewes had an IELISA-B. canis mean (%P) value of 16.18 (S.D. 5.63), while the IELISA-B. ovis had a mean (%P) value of 12.52 (S.D. 4.94). A cut-off value of (%P) 27.44 (+2 S.D.) or (%P) 33 (+3 S.D.) was determined by milk-ELISA-B. canis and (%P) 22.4 (+2 S.D.) and (%P) 27.34 (+3 S.D.) by milk-IELISA-B. ovis. These cut-off values were adjusted by receiver-operator characteristics (ROC) analysis using 23 positive samples from infected ewes, which indicated a milk-IELISA-B. canis cut-off value of (%P) 33 and milk-IELISA-B. ovis of (%P) 26 with 100% sensitivity and specificity. Based on our results, we propose that, following a study of a larger number of samples, the milk-IELISA-B. canis could be considered a suitable test for the diagnosis of B. ovis brucellosis in lactating ewes.

Abstract: The encapsulation of a Brucella ovis extract (HS) in microparticles has been proved effective against experimental infections in mice. This work describes different strategies to increase the HS loading and prepare large batches as necessary to test this vaccine in ovine. The mixture of HS with beta-cyclodextrin was optimized in order to increase the HS loading in microparticles. On the other hand, TROMS ('Total Recirculation One-Machine System') led microparticles with a more homogeneous size than the laboratory or standard procedure. Moreover, the initial burst release of HS from the standard microparticles was higher than for the TROMS ones. In fact, standard microparticles displayed a higher amount of adsorbed HS. On the contrary, both preparative methods were found effective to preserve the integrity and anti-genicity of the loaded HS. In summary, beta-CD can be used to increase the loading of large hydrophobic materials and TROMS is a valid large production of antigen-loaded microparticles.

Abstract: OBJECTIVES: Treatment of human brucellosis demands antibiotic targeting into the mononuclear-phagocytic system. The aim of this work was to prepare and characterize particulate carriers containing gentamicin and to study their interactions with phagocytic cells and bactericidal activity against intracellular Brucella melitensis. METHODS: Different poly(lactide-co-glycolide) (PLGA) polymers with free carboxylic end-group were used to formulate micro- and nanoparticles containing gentamicin, by a water-oil-water solvent-evaporation technique. PLGA 502H and 75:25H microparticles were selected because they showed the highest gentamicin loadings as well as good physico-chemical properties and sustained release in vitro. RESULTS: Gentamicin-containing microspheres of both polymers were successfully phagocytosed by infected THP-1 human monocytes, and immunocytochemistry studies revealed that the antibiotic reached Brucella-specific compartments. A dose of 30 microg of encapsulated gentamicin was able to reduce intracellular Brucella infection by 2.2 log. CONCLUSIONS: Altogether, these results suggest that 502H and 75:25H microspheres are suitable carriers for gentamicin targeting inside human macrophages and thus for brucellosis treatment.

Abstract: The recent description of the complete genomes of the two most pathogenic species of Brucella opens the way for genome-based analysis of the antigenicity of their proteins. In the present report, we describe a bench-level high-efficiency cloning and expression system (HECES) that allow expression of large numbers of Brucella proteins based on genomic sequence information. Purified proteins are produced with high efficiency in a microarray format conducive to analysis of their sero-reactivity against serum from immunized animals. This method is applicable at either small or large scale of protein processing. While it does not require robotics, the format is amenable to robotic implementation for all aspects of the process and subsequent analysis of protein characteristics. This method will allow selection of new reagents for diagnosis of brucellosis and development of vaccine against Brucella, an important zoonotic disease and biothreat agent.

Abstract: Leptospirosis, brucellosis and toxoplasmosis are widely-distributed zoonosis, being the man an accidental participant of their epidemiological chains. The aim of this paper was to make a seroepidemiological report and identify occupational and environmental variables related to these illnesses in 150 workers in a slaughterhouse in the Northern region of Paraná. For the diagnosis of leptospirosis a microscopical seroagglutination test was applied; for brucellosis, the tamponated acidified antigen test and the 2-mercaptoetanol tests were used, and for toxoplasmosis the indirect immunofluorescence reaction test. For each employee an epidemiological survey was filled, which investigated occupational and environmental variables which could be associated with these infections. Positive results for leptospirosis were found in 4.00% of the samples, for brucellosis in 0.66% of samples and toxoplasmosis in 70.00%. From the three diseases researched, only the results for leptospirosis suggest occupational infection.

Abstract: Two Gram-negative, rod-shaped, oxidase-positive, non-spore-forming, non-motile bacteria (CCUG 46016(T) and CCUG 33852(T)), isolated from a knee aspirate of a 66-year-old man and an industrial glue, respectively, were studied for their taxonomic position. On the basis of chemotaxonomic data [i.e. major ubiquinone (Q-10), major polar lipids (phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine) and major fatty acids (C(18 : 1)omega7c and C(19 : 0) cyclo omega8c)] and 16S rRNA gene sequence similarity, both strains belong to the Alphaproteobacteria. The presence of spermidine and putrescine as the predominant polyamines in CCUG 46016(T) were in agreement with its phylogenetic affiliation in the vicinity of the genus Ochrobactrum. 16S rRNA gene sequence similarities between both strains and established species within the genera Bartonella, Ochrobactrum and Brucella were less than 95 %. Although both organisms showed highest 16S rRNA gene sequence similarity to members of Brucella, phenotypic features (including chemotaxonomic features) were more like those of members of the genus Ochrobactrum. Sequence comparison of the recA genes confirmed the separate phylogenetic position of the two strains. On the basis of DNA-DNA pairing results and physiological and biochemical data, the two strains can be clearly differentiated from each other and from all known Ochrobactrum species. It is evident that these organisms represent two novel species in a new genus, Pseudochrobactrum gen. nov., for which the names Pseudochrobactrum asaccharolyticum sp. nov. (the type species, type strain CCUG 46016(T)=CIP 108977(T)) and Pseudochrobactrum saccharolyticum sp. nov. (type strain CCUG 33852(T)=CIP 108976(T)) are proposed.

Abstract: The current study was carried out to assess the use of immunomagnetic separation-polymerase chain reaction (IMS-PCR) in direct detection of Brucella abortus and B. melitensis from soft cheese and to examine a relatively small number of field samples for the presence of these species. Two methodologies, one with IMS and the other without IMS, were employed for recovery of the Brucella species from cheese samples. IMS in conjunction with the PCR assay was determined to detect as low as 3x10(2) bacteria/mL, while the limit of detection with the other extraction procedure was 3x10(3) bacteria/mL. In the analysis of 40 cheese samples collected from various markets, only B. abortus was detected by PCR using both DNA extraction procedures in two (5%) samples. No positive results were obtained by culture and B. melitensis was not found in any cheese samples examined. The results suggest that this technique is promising owing to its pace and high sensitivity and should aid in direct detection of Brucella species from complex food samples.

Abstract: A cross-sectional study was performed in the livestock-wildlife interface areas of Lochinvar and Blue Lagoon National Parks and the non-interface area of Kazungula to determine the prevalence of antibodies to Brucella spp. in domestic ruminants and identify individual animal risk factors of infection. A total of 1245 cattle from 124 herds and 280 goats and sheep from 29 flocks were tested sequentially for Brucella antibodies using the Rose Bengal test (RBT) and competitive ELISA. In cattle, individual seroprevalence ranged from 14.1% to 28.1%, while herd sero-prevalence ranged from 46.2% to 74.0% in the three study areas. No goat or sheep tested positive for Brucella antibodies. Three types of cattle grazing strategies were encountered: locally grazed herds (LGH), transhumantly grazed herds (TGH) and river flood plain grazed herds (FGH). Brucella seroprevalence was seen to vary according to area and grazing strategy: Lochinvar and transhumant grazed herds recorded the highest figures, respectively. Age, sex and history of abortion were found to have independent effects on individual seroprevalence. This study establishes that brucellosis is endemic in domestic animals in the livestock-wildlife interface areas of Blue Lagoon and Lochinvar national parks and the disease is also present in Kazungula. We observed that type of grazing strategy had significant impact on cattle Brucella seroprevalence and that transhumant herds were at high risk of being infected.

Abstract: The aim of this study was to evaluate the brucellosis patients inhabiting in Mus province, in Eastern Anatolia of Turkey, retrospectively. The mean age of the patients (n: 87) was 38.1 +/- 12.4 years, and 45% of them were female. The transmission route was the consumption of unpasteurized fresh cheese (in 85%), and unboiled milk (in 45%). The most common symptoms were recorded as chills (89%), fever (87%), and arthralgia (81%). Splenomegaly (71%) and hepatomegaly (63%) were the predominant physical examination signs. Diagnosis was made based on the clinical features and positive Rose-Bengal test result (93%), however, blood cultures could not be performed due to insufficient laboratory equipment. In 92% of the patients at least one complication has been detected indicating delayed admission to the hospital, while the most common complications were sacroileitis (79%) and spondylitis (44%). Streptomycin+doxycyclin, streptomycin+doxycyclin+ ciprofloxacin, and streptomycin+doxycyclin+ rifampicin combination therapies were used in 62%, 24% and 14% of the patients, respectively, for six weeks, resulting with complete cure.

Abstract: BACKGROUND: Brucellosis is an endemic disease present in many countries worldwide, but it is rare in Europe and North America. Nevertheless brucella is included in the bacteria potentially used for bioterrorism. The aim of this study was the investigation of the antibiotic susceptibility profile of brucella isolates from areas of the eastern Mediterranean where it has been endemic. METHODS: The susceptibilities of 74 Brucella melitensis isolates derived from clinical samples (57) and animal products (17) were tested in vitro. The strains originate from Crete (59), Cyprus (10), and Syria (5). MICs of tetracycline, rifampicin, streptomycin, gentamicin, norfloxacin, ciprofloxacin, levofloxacin, trimethoprim/sulfamethoxazole, ampicillin, amoxicillin/clavulanic acid, and erythromycin were detected by E-test method. The NCCLS criteria for slow growing bacteria were considered to interpret the results. RESULTS: All the isolates were susceptible to tetracycline, streptomycin, gentamicin, ciprofloxacin, norfloxacin, and levofloxacin. Two isolates presented reduced susceptibility to rifampicin (MIC value: 1.5 mg/l) and eight to SXT (MIC values: 0.75-1.5 mg/l). Erythromycin had the highest (4 mg/l) MIC90value and both norfloxacin and erythromycin the highest (1.5 mg/l) MIC50 value. CONCLUSION: Brucella isolates remain susceptible in vitro to most antibiotics used for treatment of brucellosis. The establishment of a standardized antibiotic susceptibility method for Brucella spp would be useful for resistance determination in these bacteria and possible evaluation of bioterorism risks.

Abstract: A serological study was carried out in Tiaret province in western Algeria on 1032 cows distributed in 95 flocks to estimate the prevalence of Brucella infection and to compare the sensitivity and specificity of a range of agglutination tests. Screening tests showed 31.5% of herds positive using the buffered plate antigen test and 26.3% using the rose Bengal test compared with 15.7% with the complement fixation test. Using the complement fixation test as the gold standard for confirmatory tests, the Rivanol test was found to be more sensitive but less specific than tube agglutination in detecting brucellosis infection. Three isolates were identified from 105 blood samples from humans with brucellosis and 50 samples of milk and tissues from infected cows and they were all Brucella melitensis biovar 3.

Abstract: A young infant with acute brucellosis is reported. He presented with a septicaemia-like picture. Diagnosis was based on a fourfold rise of Brucella agglutination titres and a positive blood culture. He had been exclusively breast-fed when his mother developed brucellosis 4 weeks after delivery. It is strongly suspected that the transmission of Brucella melitensis to this infant was through the maternal breast-milk.

Abstract: A cross-sectional study was conducted to determinate the seroprevalence rate of equine brucellosis in the state of Tamaulipas, Mexico. Serum samples from 420 equines were analyzed with the Rose Bengal test at cell concentrations of 3% (RBT-3%) and 8% (RBT-8%), and positive results were confirmed with the Rivanol test (RT). Risk factors were determined with the prevalence ratio (PR) and the use of variables generated from a questionnaire administered to the animals' owners. Serum from 1 stallion had positive results with both the RBT-8% and the RT, for a seroprevalence rate of 0.238%. Drinking of water from a pond that was also used by cattle and dogs was the only associated risk factor for this animal (PR = 0.25). However, the results were considered false-positive, because the results for other horses in the same environmental conditions were negative. Although brucellosis is considered endemic in ruminants in the study area, the results obtained suggest that equines are not a reservoir of brucellosis and do not play an important role in the epidemiologic patterns of this disease in northeastern Mexico.

Abstract: Brucellosis is a worldwide zoonosis caused by different species of the genus Brucella. The intracellular localisation of this pathogen, particularly in macrophages, renders treatment difficult since most antibiotics known to be efficient in vitro do not actively pass through cellular membranes. As alternative to current treatment, polymeric drug delivery systems containing gentamicin have been developed. These particulate carriers target the drug into the mononuclear-phagocytic system, where the pathogen resides that will allow intracellular accumulation of the antibiotic after particle degradation. Besides, particle uptake may induce macrophage activation, increasing the production of reactive oxygen intermediates, involved in host defense against the intracellular pathogen. The aim of the present work was to study the suitability of polymeric nanoparticles for gentamicin entrapment in view to treat brucellosis. Different poly(lactide-co-glycolide) PLGA polymers were used to formulate the nanoparticles containing gentamicin by a water-oil-water solvent evaporation method. Furthermore, in vitro macrophage activation upon nanoparticles phagocytosis and in vivo distribution of the nanocarriers in the target organs for Brucella (liver and spleen) were also studied. The nanoparticle sizes were below 350 nm, the gentamicin encapsulation efficiency depended on the polymer type used for their preparation and the in vitro release of the antibiotic exhibited a continuos pattern (PLGA 502H). PLGA 502H nanoparticles were the most suitable due to the highest entrapment and the most sustained release. The nanoparticles were successfully phagocyted by a J774 murine monocytes cell line and biodistribution studies in mice after intravenous administration of the delivery systems revealed that the particles reached the target organs of Brucella (liver and spleen). All together, these results indicate that the nanocarriers described in this work may be suitable as gentamicin delivery system to control brucellosis.

Abstract: Forty weaned male guinea pigs of 208.20 +/- 6.62 g mean body weight were divided into 4 groups of 10 animals in a randomized block design. All of the guinea pigs were fed a basal diet [25% ground maize hay, 30% ground maize grain, 22% ground chickpea (Cicer arietinum L.), 9.5% deoiled rice bran, 6% soybean meal, 6% fish meal, 1.45% mineral supplement (without Zn) and 0.05% ascorbic acid] and available green fodder. Group I served as the control (no Zn supplementation), whereas 20 ppm Zn was added in the diet in groups II, III, and IV either as zinc sulfate (ZnSO(4)), zinc amino acid complex (ZAAC), and ZnSO4 + ZAAC in equal parts, respectively. Experimental feeding lasted for 70 d, including a 3-d digestibility trial. Blood was collected through cardiac puncture from four animals in each group at d 0 and subsequently at the end of experimental feeding. After 40 d of experimental feeding, four animals from each group were injected with 0.4 mL of Brucella abortus cotton strain-19 vaccine to assess the humoral immune response of the animals. After 10 wk of study, four animals from each group were sacrificed to study the concentration of Zn, Cu, Co, Fe, and Mn in the liver, pancreas and spleen. Results revealed no significant difference in the feed intake, body weight gain, and digestibility of the nutrients, except for crude protein (CP) digestibility, which was significantly (p < 0.05) lower in group IV. Although concentrations of serum glucose, Ca, and P and the albumin:globulin (A:G) ratio were similar in the different groups, the total protein, albumin, and serum alkaline phosphatase activity were higher in all of the Zn-supplemented groups on d 70. The serum Zn levels at the end of experimental feeding were significantly higher in groups II and III, whereas serum Mn levels were found to be significantly (p < 0.05) higher in groups III and IV. The organ weights (as percentage of body weights) did not show any differences among the treatment groups. Although the Mn concentration was significantly (p < 0.05) higher in the pancreas, the Cu concentration was significantly (p < 0.05) reduced in the spleen in all of the Zn-supplemented groups. The humoral immune response (antibody titer values) on d 14 of vaccination was significantly (p < 0.05) higher in all of the Zn-supplemented groups. It was concluded that the 20-ppm level of Zn in the diet might be adequate for growth and nutrient utilization in guinea pigs, but supplementation of 20-ppm zinc significantly improved the immune response and impact was more prominent with the ZAAC (organic source) compared to ZnSO(4) (inorganic source).

Abstract: In countries such as Mexico, brucellosis is still an important public health problem due to the consumption of non-pasteurized milk and dairy products, contaminated with Brucella spp. The aim of this study was to look into the survival of Brucella abortus during fermentation of milk with a yoghurt starter culture and storage at refrigeration temperature. Sterile skim milk was inoculated with B. abortus at two concentrations, 10(5) and 10(8) CFU/ml simultaneously with a yoghurt starter culture of lactic acid bacteria (Streptococcus thermophilus and Lactobacillus delbrueckii subspecie bulgaricus). Inoculated flasks were incubated at 42 degrees C, followed by refrigeration at 4 degrees C. Samples were taken during fermentation and during storage and viable count of B. abortus and lactic acid bacteria and pH were determined. Results showed that after 10 days of storage at 4 degrees C, B. abortus was recovered in fermented milk at a level of 10(5) CFU/ml, despite the low pH below 4.0. Therefore B. abortus is able to survive in fermented milk. This finding may imply that non-pasteurized fermented milk contaminated with Brucella abortus could be a means of transmission of these bacteria.

Abstract: The commonly used live attenuated vaccine in ovine brucellosis prophylaxis is Brucella melitensis Rev.1. This vaccine is known to induce antibody responses in vaccinated animals indistinguishable by the current conventional serological tests from those observed in challenged animals. Brucella BP26 and Omp31 proteins have shown an interesting potential as diagnostic antigens for ovine brucellosis. Accordingly, the bp26 gene and both bp26 and omp31 genes have been deleted from the vaccine strain Rev.1. Immunogenicity and vaccine efficacy of the parental Rev.1 strain and of both mutants in protecting sheep against B. melitensis strain H38 challenge was evaluated by clinical and bacteriological examination of ewes. They were conjunctivally or subcutaneously vaccinated when 4 months old and then challenged with B. melitensis H38 at the middle of the first pregnancy following vaccination. Deletion of bp26 and omp31 genes did not significantly affect the well recognised capacity of Rev.1 to protect sheep against B. melitensis challenge. However, the protection conferred by the CGV2631 mutant was significantly lower than that conferred by the CGV26 mutant or the Rev.1 strain. Vaccinated and challenged animals were detected positive in classical serological tests and in the IFN-gamma assay. A BP26-based ELISA was investigated to discriminate between ewes vaccinated by the mutants and ewes challenged with B. melitensis H38. The cut-off which was chosen in order to have 100% specificity resulted in a moderate sensitivity for the detection of challenged ewes. The use in the field of one of the mutants as vaccine against a B. melitensis infection, combined with classic diagnostic tests and a BP26 ELISA, could thus give an improvement in the differentiation between vaccinated and infected animals and contribute to the objective of eradication of brucellosis in small ruminants.

Abstract: Food safety was identified as a high priority area in the 2001-2005 World Organisation for Animal Health (OIE) Strategic Plan. Member Countries of the OIE considered that the organisation should be more active in issues of public health and consumer protection and that this should include more involvement in the area of diseases or pathogens transmissible through food, whether or not animals are affected by such diseases or pathogens. A permanent Working Group on Animal Production Food Safety was established in 2002 to coordinate the OIE's activities in food safety. The Working Group was requested to focus on food safety measures applicable at farm level and to monitor the ongoing cooperation between the OIE and Codex Alimentarius. More emphasis is now placed on the public health aspects of a disease when OIE standards are developed or revised. For example, the revised chapter on bovine tuberculosis in the Terrestrial Animal Health Code includes food safety recommendations for meat and meat products and for milk and milk products. The revised chapter was approved by the OIE International Committee of Member Countries at their 73rd General Session in May 2005. More chapters will follow, beginning with a chapter addressing bovine brucellosis.

Zakhlebnaia OD, Laukner IV. [The use of polyvinylpyrrolidone as a stabilizer in the lyophilization of Brucella]. Laboratornoe delo. 1991; (2); 62-3. [PubMed: 1709995].

Abstract: Stabilizing effects of 5 percent aqueous polyvinylpyrrolidone (PVP) solution and sucrose-agar-gelatin medium (SAG) were examined in lyophilization of 4 Brucella types. The number of viable cells immediately after lyophilization in PVP has made up 48.1 percent on an average; this value was different with different Brucella species: 70.9 percent with B. abortus, 50.4 percent with B. melitensis, 41.2 percent with B. rangiferi, 30.2 percent with B. suis. Only 27.5 percent of cells were viable after Brucella lyophilization in SAG medium. Gradual death of lyophilized cells was observed in the course of strain storage at 5 degrees C. After two-year storage the number of viable cells after lyophilization in PVP was 41.8, after three-year storage 35.1 percent. The number of viable cells after SAG lyophilization was still lower. Biological characteristics of the strains lyophilized in both media corresponded to the initial ones over the three-year follow-up.

Abstract: Polyhydroxyalkanoates (PHAs) are gaining more and more importance the world over due to their structural diversity and close analogy to plastics. Their biodegradability makes them extremely desirable substitutes for synthetic plastics. PHAs are produced in organisms under certain stress conditions. Here, we investigated 253 sequenced (completely and unfinished) genomes for the diversity and phylogenetics of the PHA biosynthesis. Discrepancies in the phylogenetic trees for phaA, phaB and phaC genes of the PHA biosynthesis have led to the suggestion that horizontal gene transfer (HGT) may be a major contributor for its evolution. Twenty four organisms belonging to diverse taxa were found to be involved in HGT. Among these, Bacillus cereus ATCC 14579 and Xanthomonas axonopodis pv. citri str. 306 seem to have acquired all the three genes through HGT events and have not been characterized so far as PHA producers. This study also revealed certain potential organisms such as Streptomyces coelicolor A3(2), Staphylococcus epidermidis ATCC 12228, Brucella suis 1330, Burkholderia sp., DSMZ 9242 and Leptospira interrogans serovar lai str. 56601, which can be transformed into novel PHA producers through recombinant DNA technology.

Abstract: A competitive ELISA (Brucella-Ab c-ELISA) was standardized and validated for the detection of Brucella antibodies in cattle, sheep and goat sera using a monoclonal antibody (MAb 4B5A) produced against Brucella melitensis biotype 2. The specificity and sensitivity of the assay were 100% to a 67.5% cut-off point (B/Bo%). When compared with an indirect ELISA, the Brucella-Ab c-ELISA did not demonstrate cross-reactions when testing positive sera for antibodies to some Enterobacteriaceae. A comparison was made between the Brucella-Ab c-ELISA and the complement fixation and Rose Bengal tests. Results demonstrated that the Brucella-Ab c-ELISA is a valuable tool for the serological diagnosis of bovine and ovine/caprine brucellosis.

Abstract: The aim of this study was to evaluate the activity of gamma interferon (IFN-gamma) when it was either adsorbed onto or loaded into albumin nanoparticles. Brucella abortus-infected macrophages and infected BALB/c mice were selected as the models for testing of the therapeutic potentials of these cytokine delivery systems, in view of the well-established role of IFN-gamma-activated macrophages for the control of Brucella sp. infections. Whereas the encapsulation of IFN-gamma inside the matrix of nanoparticles completely abrogated its activity, adsorbed IFN-gamma increased by 0.75 log unit the bactericidal effect induced by RAW macrophages activated with free IFN-gamma, along with a higher level of production of nitric oxide. In infected BALB/c-mice, IFN-gamma adsorbed onto nanoparticles was also more active than free cytokine in reducing the number of bacteria in the spleens, and the effect was mediated by an increased ratio of IFN-gamma-secreting (Th1) to interleukin-4-secreting (Th2) cells. Overall, albumin nanoparticles would be suitable as carriers that target IFN-gamma to macrophages and, thus, potentiate their therapeutic activity.

Abstract: Drug delivery systems containing gentamicin were studied as a treatment against experimental brucellosis in mice. Micro- and nanoparticles prepared by using poly(D,L-lactide-coglycolide) (PLGA) 502H and microparticles made of PLGA 75:25H were successfully delivered to the liver and the spleen, the target organs for Brucella melitensis. Both polymers have the same molecular weight but have different lactic acid/glycolic acid ratios. Microparticles of PLGA 502H and 75:25H released their contents in a sustained manner, in contrast to PLGA 502H nanoparticles, which were degraded almost completely during the first week postadministration. The values of the pharmacokinetic parameters after administration of a single intravenous dose of 1.5 mg/kg of body weight of loaded gentamicin revealed higher areas under the curve (AUCs) for the liver and the spleen and increased mean retention times (MRTs) compared to those for the free drug, indicating the successful uptake by phagocytic cells in both organs and the controlled release of the antibiotic. Both gentamicin-loaded PLGA 502H and 75:25H microparticles presented similar pharmacokinetic parameter values for the liver, but those made of PLGA 75:25 H were more effective in targeting the antibiotic to the spleen (higher AUCs and MRTs). The administration of three doses of 1.5 mg/kg significantly reduced the load associated with the splenic B. melitensis infection. Thus, the formulation made with the 75:25H polymer was more effective than that made with 502H microspheres (1.45-log and 0.45-log reductions, respectively, at 3 weeks posttreatment). Therefore, both, pharmacokinetic and pharmacodynamic parameters showed the suitability of 75:25H microspheres to reduce the infection of experimentally infected mice with B. melitensis.

Abstract: Brucellosis is a rarely encountered zoonosis in Poland caused by Brucella species. Most of cases are imported from endemic areas. We reported 2 cases (married couple) oftypial brucellosis in patient returning from Azerbaijan, where both of them drunk non-pasteurized goat milk. The diagnosis was established by serological tests. Brucellosis should be suspected in patients with unexplained fever, especially in travelers to countries where non-pasteurized dairy products are common.

Abstract: Brucella abortus, a facultative intracellular pathogen, is of tremendous zoonotic importance because of its ability to induce spontaneous abortion in cattle and other livestock. It is also known to cause persistent undulant fever, endocarditis, arthritis, osteomyelitis and meningitis in humans. The available vaccines against this dreadful infection suffer from limitations like short-term immunity, increased risk of hypersensitivity and low prophylactic index in the recipients. In the present study, we have demonstrated that liposomal form of a recombinant ribosomal L7/L12 protein, a B-T cell antigen of B. abortus, activates strong immune response in the host. In contrast, free antigen generates moderate immune response in the immunised animals. The liposomisation of rL7/L12 protein causes tremendous increase in cell-mediated immune response in terms of delayed type hypersensitivity, T-cell proliferation and up-regulation in type I cytokine expression, etc. Moreover, the liposome encapsulated antigen elicited stronger humoral immune response as compared to standard vaccine (S-19) or IFA-L7/L12 combination in the immunised animals. The effectiveness of liposome-based vaccine was also substantiated by better systemic clearance of bacterial load after challenging the animals with B. abortus 544 pathogen. The results of the present study suggest the potential of liposome-based rL7/L12 antigen as prospective and efficient candidate vaccine capable of eliciting both cell mediated as well as humoral immune responses against experimental murine brucellosis.

Abstract: The principal methods for the serological diagnosis of bovine brucellosis are the complement fixation test (CFT), serum agglutination test (SAT), Rose-Bengal test (RBT), indirect enzyme-linked immunosorbent assay (iELISA) and more recently the competitive ELISA (cELISA) and the fluorescent polarisation assay (FPA). Guidelines set by the World Organisation for Animal Health (OIE) describe methods and diagnostic thresholds for each of these tests. Many countries have adopted these methods for the purposes of eradication of brucellosis and have legislated for the use of these tests (the CFT and SAT in particular) for the prevention of the spread of the disease through international trade. Within the European Union (EU) each member state has a National Reference Laboratory which regulates the quality of brucellosis diagnosis and works to the recommendations set by the OIE. This article describes the results from the first three EU ring trials assessing the harmonisation of diagnostic tests between each member state. The general level of harmony for SAT, CFT, and iELISA was found to be good, but issues of standardisation of the RBT, cELISA and FPA remain. The cELISA and FPA in particular need further work to create European harmony. The ring trials also proved successful at providing specific evidence of poor performance in some areas. The decision on whether or not to take action on the basis of these results rested with the individual laboratories concerned. The increase in the number of participants in these trials over time reflected the enlargement of the EU and increased the need for quality assurance.

Abstract: Brucellosis is a zoonotic disease and virtually all infections derived from exposure to animals or ingestion of unpasteurized dairy products. Brucellosis among family members has been reported. However, screening household members of an index case of acute brucellosis is not a routine procedure. A 10-y-old boy was diagnosed with acute brucellosis. Unpasteurized goat cheese commonly consumed within the family was thought to be the possible source of the bacteria. The family (parents, sister and brother) was screened with physical examination, serum tube agglutination test, blood cultures and routine laboratory tests. Three additional cases (parents and sister) of serological and culture proven brucellosis were detected. Two of them (mother and sister) were asymptomatic and had no clinical findings. Brucella melitensis biovar 3 was isolated from breast milk culture and from all blood cultures of 4 brucellosis cases. In conclusion, brucellosis, even with bacteraemia, can be completely asymptomatic. Consumption of raw milk products by household members is a common risk factor for brucellosis outbreak among family members. Thus, screening household members of an index case of brucellosis can expose new brucellosis cases.

Abstract: Between 1987 and 1990, three patients with abscesses produced by Brucella endocarditis were admitted to the Department of Infectious Diseases, "Ospedali Riuniti", Bergamo, Italy. In each case, the diagnosis was based on a history of ingestion of milk products, positive Wright serology, positive blood and valvar culture, and echocardiography. Medical therapy alone was not found to be effective in treatment, all patients requiring surgical intervention. One case required urgent surgical treatment and underwent three further operations up to the final implantation of a valved tube. According to our experience, Brucella endocarditis is a rare but serious disease which requires a combination of medical and surgical therapy.

Abstract: OBJECTIVE: Brucellosis is a Zoonosis, recognized worldwide as a serious public health hazard and economically significant disease. The etiologic agent of this disease is a bacterial species of the genus Brucella that causes undulant fever, sterility and other systemic illnesses in human. Thus, the aim of this study was to undertake seroepidemiological study of brucellosis on occupationally exposed persons in Addis Ababa abattoirs enterprise and different dairy farms. METHODS: Three hundred thirty six human individuals were screened using Rose Bengal Plate Test (RBPT) and the sera were further subjected to 2- Mercapto Ethanol Test (2-MET). Simultaneously, survey was conducted in farms and slaughterhouse to investigate epidemiological factors. RESULTS: An overall seroprevalence rate of 4.8% (16/336) was determined by taking RBPT+/2MET+ as confirmatory test (P < 0.05). Some of the risk factors contributing to the occurrence of the disease include occupation, gender, and unsafe handling of infected materials. Raw milk and meat consumption, lack of awareness and use of detergents were also found as important factors. CONCLUSION: The seroprevalence of brucellosis is expected to be more in other areas of the Region, where there is high human-animal contact and high number of cattle population with a respective traditional system of animal rearing.

Abstract: We report five cases with unusual causes of intrahepatic cholestasis, including consumption of Teucrium polium (family Lamiaceae) in the form of tea, Stauffer's syndrome, treatment with tamoxifen citrate for breast cancer, infection with Coxiella Burnetii (acute Q fever), and infection with Brucella melitensis (acute brucellosis).

Abstract: Brucella spp. is the causative agent of brucellosis (Malta fever), which is the most widespread zoonosis worldwide. The pathogen is capable of establishing persistent infections in humans which are extremely difficult to eradicate even with antibiotic therapy. Moreover, Brucella is considered as a potential bioterrorism agent. Histidinol dehydrogenase (HDH, EC 1.1.1.23) has been shown to be essential for the intramacrophagic replication of this pathogen. It therefore constitutes an original and novel target for the development of anti-Brucella agents. In this work, we cloned and overexpressed the HDH-encoding gene from Brucella suis, purified the protein and evidenced its biological activity. We then investigated the inhibitory effects of a series of substituted benzylic ketones derived from histidine. Most of the compounds reported here inhibited B. suis HDH in the lower nanomolar range and constitute attractive candidates for the development of novel anti-Brucella agents.

Abstract: Helicobacter pylori infection is an important risk factor for gastric diseases. Some probiotics are useful for suppressing H. pylori infection. Bifidobacterium bifidum YIT 4007 can improve the experimental gastric injury in rats and the disease stages on the gastric mucosa in peptic ulcer patients. We evaluated the fermented milk using a clone (BF-1) having the stronger ability to survive in the product than this parent strain to clarify the in vitro suppressive effect of BF-1 on H. pylori and the in vivo efficacy of BF-1 fermented milk on H. pylori and gastric health. In the mixed culture assay of BF-1 and H. pylori, the number of pathogens was decreased such that it was not detected after 48 h in the Brucella broth with a decrease in pH values. In the cell culture experiment with human gastric cells, the H. pylori infection-induced IL-8 secretion was suppressed by the preincubation of BF-1. In a human study of 12-wk ingestion (BF-1 group, n = 40; placebo group, n = 39) with a randomized double-blind placebo-control design, the H. pylori urease activity and gastric situation were evaluated using a urea breath test (UBT) and the serum pepsinogen (PG) levels as biomarkers for inflammation or atrophy, respectively. In the H. pylori-positive subjects, the difference (DeltaUBT) of the UBT value from the baseline value in the BF-1 group (n = 34) was lower than that in the placebo group (n = 35) at 8 wk. The baseline UBT values showed a negative correlation with DeltaUBT values at 8 and 12 wk in the BF-1 group but not in the placebo. In the PG-positive subjects classified by the PG test method, the BF-1 group was lower in DeltaUBT values than the placebo group at 8 and 12 wk. In the active gastritis class by PG levels, the BF-1 group was lower in their DeltaUBT values than the placebo at 8 and 12 wk. The PG I levels in the BF-1 group were lower than the placebo at 12 wk. The PG II levels in the BF-1 group did not change during the ingestion period, but the placebo was increased. The PG I/II ratios slightly decreased from baseline at 12 and 20 wk in the BF-1 and placebo groups. These patterns were also observed in the H. pylori-positive subjects. The improving rates of upper gastrointestinal symptomatic subjects and total symptom numbers in the BF-1 group were higher than those in the placebo. These results indicate that BF-1 fermented milk may affect H. pylori infection or its activity, gastric mucosal situation, and the emergence of upper gastrointestinal symptoms.

Abstract: A young male immigrant from Syria with a vasculitic-appearing leg rash, asymmetrical polyarthritis, microscopic haematuria, and raised inflammatory markers was provisionally diagnosed with Henoch-Schönlein purpura. Skin biopsy showed leukocytoclastic vasculitis. Low-grade fevers persisted despite non-steroidal anti-inflammatory therapy, and Brucella sp. was subsequently grown from both blood and synovial fluid aspirates. Further tests gave positive results for B. abortus, and triple antibiotic therapy produced a rapid clinical response. Cutaneous vasculitis has rarely been described in brucellosis, and this is the first report in the English medical literature of brucellosis mimicking Henoch-Schönlein purpura.

Abstract: Minimal nutritional requirements and temperature limits of growth were studied in Brucella suis and, comparatively, in a few other Brucella species. In a saline basic medium including thiosulphate, ammonium sulphate and glucose with addition of 2 or 4 vitamins (nicotinic acid, thiamin and panthotenic acid, biotin), 24 out of 25 B. suis, 4/6 B. melitensis and 1/6 B. abortus strains were able to grow. Some strains, however, needed to be initially induced to grow by other ingredients, CO2, other vitamins, or amino acids, or by a prolonged incubation. In the saline basic medium without ammonium, glutamic acid and/or alanine and arginine, with or without glucose, supported the growth of all the B. suis and B. melitensis strains, except 2 which required a sulphur amino acid. Five out of 6 B. abortus strains did not grow in either medium without addition of one or several aromatic amino acids or, for one strain, aspartic acid, or valine. One strain could also be induced to grow in ammonium medium by other amino acids. In a rich medium with yeast extract, all Brucella species grew at 18 degrees C and 42.5 degrees (except one) while most B. suis (14/17) grew also at 15 degrees C and 44 degrees C, in contrast to other brucellae of which a few strains only grew at these temperatures. In saline ammonium glucose medium, yeast extract at 0.1 g/l provided all the required vitamins and amino acids for all brucellae and at 1 g/l, it even provided enough nitrogen to support growth without ammonium. Such basic saline medium with yeast extract may be advantageously used in routine Brucella culture, instead of the classic undefined peptone mediums. B. suis biovar 1 strains did not differ significantly in their minimal nutritional requirements, precluding the use of these requirements to differentiate the strains, in particular the Chinese vaccine strain S2 from the reference strain 1330 or from other strains from different parts of the world. Finally, B. suis which is endowed with a nearly complete synthetic potential may represent the parental Brucella species from which the melitensis and abortus species may have evolved.

Abstract: Activation of macrophages and their antimicrobial activities by interferon-gamma (IFN-gamma) plays a crucial role in controlling Brucella infection. Interleukin-4 (IL-4) antagonizes the macrophage activity effects of IFN-gamma and thus inhibits cell-mediated immune reactions. Given that the production of IFN-gamma and IL-4 are under genetic control, we investigated the relationship between these two cytokine gene polymorphisms and the susceptibility to brucellosis. Hundred and ninety-five patients with brucellosis and 91 healthy animal husbandmen who owned infected animals and consumed their contaminated dairy products were selected to participate in this study. All individuals were genotyped for IFN-gamma and IL-4 gene polymorphisms at positions +874 and -590, respectively. Results showed that IFN-gammaAA genotype was significantly more prevalent (P =0.03) and IL-4CC genotype was significantly less frequent (P =0.034) in the patient group compared to the control group. Also, the frequency of IFN-gamma/IL-4 combination of genotype (IFN-gammaTT/IL-4CC) and allele (IFN-gammaT/IL-4C) were significantly higher in the controls than in the patients (P =0.033 and P =0.0035, respectively). Data suggest that individuals who have IFN-gammaAA genotype are more susceptible, and those who carry IL-4CC genotype are more resistant to brucellosis. We also suggest that individuals who carry IFN-gammaT/IL-4C or IFN-gammaTT/IL-4CC can be more resistant to Brucella infection.

Abstract: The aim of this study was to investigate the adjuvant properties of oral-administered Gantrez AN nanoparticles with ovalbumin (as allergen model) and, in some cases, lipopolysaccharide of Brucella ovis as immunomodulator. For this purpose, BALB/c mice were administered by oral gavage with OVA nanoparticles and both Th1 and Th2 markers (IgG2a and IgG1, respectively) were enhanced. On the other hand, these carriers administered by oral route were able to protect a model of sensitized mice to ovalbumin from anaphylactic shock. These results are highly suggestive for the valuable use of Gantrez nanoparticles in oral immunotherapy with allergens.

Abstract: Dairy cattle submissions to the California Veterinary Diagnostic Laboratory System (CVDLS) were analyzed to determine submitter statistics. Eligible submissions were those received July 1, 1987 through December 31, 1989 for reasons other than regulatory brucellosis serology. A comprehensive list frame of California dairies was constructed from Brucella Ring Test information and served as the comparison population for the study. Analyses were performed based on geographic location, herd size, proximity to a CVDLS laboratory, and frequency of submission. Thirty-nine percent of the 2,490 California dairies in the reference population had submitted specimens greater than or equal to 1 time to the CVDLS during the study period. Twenty-three percent of the reference population had submitted greater than or equal to 2 times. Specimens were more likely to be submitted from larger herds than smaller herds. Larger dairies also submitted specimens more frequently. Dairies in the northern part of the state were more likely to submit specimens and submitted more frequently than southern herds when herd size was accounted for in the analysis. Mean submission rate (+/- SD) for the 970 submitting dairies was 1.4 (+/- 1.8) submissions/year. Forty-six percent of the dairies accounted for 80% of submissions, whereas only 14% of dairies accounted for 50% of all diagnostic dairy submissions.

Abstract: OBJECTIVES: Cases of human brucellosis in Korea have recently increased due to the increasing incidence of bovine brucellosis. The authors conducted this study to elucidate the status of brucellosis through seroepidemiologic study. METHODS: We selected our study population from a high risk group. We conducted a questionnaire survey and obtained blood samples to determine the seroprevalence of brucellosis antibodies for 10 days in February, 2005. The titers of brucellosis were measured by the combination of standard tube agglutination test (STA) and enzyme-linked immunosorbent assay (ELISA) test. RESULTS: Our study subjects comprised 1,075 cases: 971 livestock workers, 51 veterinarians, and 53 artificial inseminators. In the STA test, 27 cases (2.5%) had titers of greater than or equal to 1:20. Of 1,068 cases (7 cases were excluded due to previous brucellosis), 7 cases of brucellosis were diagnosed with titers of 1:160, giving a seroprevalence of brucellosis of 0.66%. The seroprevalence in the male group was 0.95%, and that of livestock workers, veterinarians, and artificial inseminators was 0.52%, 4.17%, and 0.00%, respectively. The Spearman's correlation coefficient between the positive rate of bovine brucellosis per capita and household and human brucellosis was 0.806 and 0.744, respectively. The concordance rate between the Korea National Institute of Health and the Gyeongsangbuk-do Institute of Health and Environment by the STA and ELISA tests was 94.7% and 100.0%, respectively. CONCLUSIONS: The study results indicated in higher seroprevalence rate among veterinarians than among livestock workers and artificial inseminators. Because veterinarians may be exposed to this high risk, effective working guidelines for veterinarians to guard against brucellosis must be developed. Moreover, more extensive epidemiologic research for laboratory workers and meat handlers is needed.

Abstract: OBJECTIVE: To study the clinical spectrum of brucellosis in Bikaner (Northwest India). METHODS: A total of 175 cases were diagnosed as brucellosis during the period of six year (June 1997 to May 2003). They were studied for clinical profile and treated by rifampicin and doxycyclin and additionally streptomycin for initial 14 days in patients of neurobrucellosis. These patients were followed up to 3 months. RESULTS: Patients of brucellosis presented with a wide spectrum of clinical manifestations. Out of 175 cases 155 were from rural area. Age ranged between 12-60 years (124 males, 51 females). Analysis of risk factors revealed history of raw milk ingestion (86.86%), occupational contact with animals (81.14%), handling of infected material (62.28%), household contact (16%) and 2 patients were veterinarian. Joint pain (83.43%) and fever (77.71%) were the commonest presenting feature. Sacroiliac joint was most commonly involved (46.86%). 31 cases had involvement of multiple joints. Other mode of presentation were neurobrucellosis (18.86%), manifested as polyradiculoneuropathy, myeloradiculopathy, meningoencephalopathy and polyradiculomyeloencephalopathy; predominant pulmonary involvement (4.0%) presented as bronchitis, pneumonia and pleural effusion; epididymoorchitis, infective endocarditis, nephrotic syndrome and recurrent abortion. All patients responded well to the treatment. CONCLUSION: Brucellosis is an important emerging zoonotic disease but it is often under-diagnosed due to lack of suspicion and diagnostic facilities despite the fact that cattle farming (an important high risk group) is one of the main occupation in rural area. This report should infuse the awareness about this reemerging disease specifically in high-risk group.

Abstract: Eighty-eight cattle were injected SC with 2.5 x 10(8) viable cells of Brucella abortus strain 19. All but 1 heifer became seropositive on the basis of the results of 7 brucellosis tests, and the proportion positive decreased with time. The proportion of cattle that were seropositive during a 20- to 67-week period after vaccination was as follows, in decreasing order: hemolysis-in-gel, 59%; buffered-acid plate antigen, 39%; ELISA, 16%; card, 10%; rivanol, 8%; cold complement-fixation, 7%; and automated complement-fixation, 5%. Using the serologic classification in Uniform Methods and Rules for brucellosis eradication, 7 cattle tested brucellosis-positive (2 suspects and 5 reactors). None of the 27 nonpregnant heifers tested positive. Of 18 heifers that were 84 to 135 days in gestation when vaccinated, 6 (33%) tested positive for brucellosis, compared with 0 of 13 and 1 (3%) of 30 heifers that were 11 to 78 and 145 to 253 days in gestation at vaccination, respectively (X2 = 12.07; 2 df; P less than 0.01). Neither breed (Angus, Hereford, Jersey, and Brahman) nor calf survival was related to brucellosis-positive results. Postpartum milk samples from 61 heifers and 24 tissues from 2 reactor cattle were culture-negative for B abortus.

Abstract: In the present study, the toxic and immunosuppressive effects of three isolates of Fusarium moniliforme (MRC 826, MRC 1069, and RRC 408) in male White Leghorn chickens were investigated. Chickens were fed diets containing .5, 5.0, or 25% of F. moniliforme corn cultures for 6 wk. About 30% of the chickens fed RRC 408 had leg weakness. Relative weights of bursae of Fabricius were lower in birds fed all doses of MRC 826. Birds fed 5.0 or 25% MRC 1069 had lower relative spleen weights. Thymus weights were lower in birds fed .5 and 5.0% RRC 408, but not in those fed 25%. Immunosuppressive effects were determined by measuring serum levels of primary and secondary agglutinin responses to SRBC and Brucella abortus. Birds fed 5.0 or 25% MRC 826 were immunosuppressed, as shown by low antibody titers to B. abortus in both the primary and secondary responses. Isolate MRC 1069 caused a decrease in secondary response to SRBC at 25% and B. abortus at the 5.0 and 25% of culture material. The RRC 408 isolated reduced the secondary response to SRBC at all doses and response to B. abortus at 5.0 and 25% of culture material. These results appear to be the first report that feed contaminated with F. moniliforme can produce deficiencies in the immune system of chickens.

Abstract: We analyzed clinically and epidemiologically 285 patients suffered from chronic brucellosis treated during 12 years. Especially we observed 51 persons with the infection called "brucellosis recens". Comparing both groups, we did not notice any statistically significant differences in the changes of organs and systems. This suggests greater influence of kind and frequency of exposition on pathologic changes than duration of illness.

Abstract: The previously reported sodium polyanethol sulfonate disk test for the identification of Peptostreptococcus anaerobius (Graves et al., 1974) was evaluated, with modifications. Three bands of brucella agar, three inoculum sizes, and two inoculum sources were compared. Nine stock cultures of P. anaerobius (eight normal flora isolates and ATCC 27337) and 16 fresh clinical isolates were used. All cultures of P. anaerobius showed inhibition zones of 12 to 30 mm in diameter, regardless of test conditions. Out of 103 clinical isolates of other species of anaerobic gram-positive cocci tested, only two had an inhibition zone size in this range (one P. micros of 11 studied had a zone of 12 mm and one P. prevotii of 14 studied had a zone of 16). The test had an overall accuracy of 98% in the identification of P. anaerobius from clinical specimens. Since P. anaerobius accounts for one-fifth to one-third of all anaerobic gram-positive cocci encountered in clinical specimens, this simple and rapid technique can be very useful for presumptive identification.

Abstract: To compare the effects of milk stasis and milk flow on Brucella abortus infection of the mammary gland under the same systemic conditions, primiparous goats (n = 5) were inoculated IV with B abortus on the day of parturition, and suckling by their neonates was restricted to one mammary gland. Goats were euthanatized and necropsied at 3 weeks after inoculation, and milk, mammary glands, and supramammary lymph nodes were evaluated by bacteriologic, histologic, and immunoenzymatic staining techniques. Nonnursed mammary glands had high titers of brucellae in milk, moderate interstitial mastitis, and brucellar antigen in macrophages located primarily in alveolar and ductal lumina. Brucellae often filled the macrophage cytoplasm. In contrast, nursed mammary glands had fewer brucellae in milk, minimal inflammatory changes, and no detectable brucellar antigen in histologic sections. Hyperplastic changes were only seen in supramammary lymph nodes draining nonnursed mammary glands; these contained more brucellae than lymph nodes draining nursed mammary glands. These studies show that milk stasis may be the sole cause of increased susceptibility of nonnursed mammary glands to B abortus infection.

Knosel H, Hempel E, Forschner E. [Experiences in the control of brucellosis in cattle herds in the government district of Hannover]. DTW. Deutsche tierarztliche Wochenschrift. 1991 Sep; 98(9); 356-8. [PubMed: 1954865].

Abstract: Caused by imported beef cattle new outbreaks of Brucellosis (subtype III) were observed in the government district of Hannover, which was "Brucellosis free" over a long period. With the aim to interrupt the series of infections from herd to herd it seemed to be necessary to introduce a herd screening system including frequent tests. It was not possible to screen the herds by the usual way of blood serum testing because of reasons of practicability and economy. A practicable alternative was the ELISA supported tank investigation. Four of the last six outbreaks were detected by this milk sampling. The other two infected herds were detected by only clinical symptoms because the lactating cows were not infected and had no contacts to the infected separately held heifers. The required test frequency in the dairy cattle herds could only be realized applying an ELISA supported highly sensitive tank milk test method. This method offered the chance of discovering infected herds as soon as possible and prevented greater economical losses and animal health risks. The tank milk screening of the dairy cattle herds to detect antibodies against Brucellosis (and additional EBL and IBR) has now become a standard method to continue the official status "free from..." This is a safer and a more economic alternative to the previous blood sampling of cattle older than two years with a three years interval because this method guarantees a safe information about the serological status of each cow which is now tested at least once a year, depending on the time of dry standing status.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: Biotechnological methods offer promising approaches for improved diagnostic and prophylactic purposes. The following biotechnological techniques are used in the Institute of Virology at the Hanover Veterinary School:--Production of monoclonal antibodies directed against viral and bacteria-specific antigens such as bovine virus diarrhoea virus, classical swine fever (hog cholera) virus, feline leukaemia virus, animal parvoviruses, Alphavirus, Brucella and Francisella--Establishment of improved and sensitive diagnostic enzyme immunoassays (ELISA) using monoclonal antibodies--Molecular cloning and sequencing of classical swine fever virus RNA and parvovirus DNA--Development of diagnostic hybridisation techniques (dot, slot, Southern and Northern blot, in situ, oligonucleotides)--Detection of viral genomes in tissues of infected animals--Development of synthetic oligopeptides as diagnostic antigens and as potential immunogens for vaccines. Currently available techniques used in basic research (e.g. pathogenesis studies) will be tested for their application in routine diagnosis of viral diseases, e.g. by molecular hybridisation. Some techniques need to be simplified (e.g. RNA extraction procedures) and, particularly, alternative labelling schedules must be developed (e.g. biotin or sulfone labelling instead of radionuclides).

Abstract: Data on costs associated with episodes of disease and disease prevention, including expenditures for veterinary services, were collected from 57 California beef cow-calf herds during 1988-1989 as part of the National Animal Health Monitoring System. Mean cost associated with episodes of disease was $33.90/cow-year, with $0.78 and $1.37/cow-year being spent for veterinary services and drugs, respectively. The highest cost for veterinary services related to episodes of disease were for dystocia, lameness, and ocular carcinoma. For disease prevention, mean expenditures for veterinary services were $1.67/cow-year, nearly all of which was spent on prevention of reproductive tract conditions. Preventive expenditures for veterinary services related to female infertility (pregnancy examination), vaccination against brucellosis and male infertility (breeding soundness examination) were $0.72, $0.39, and $0.22/cow-year, respectively. Many costs associated with episodes of disease and disease prevention were similar to those reported from Colorado National Animal Health Monitoring System beef herds.

Abstract: The milk pools of two cattle herds in Lower Saxony were found to give positive results in an enzyme-linked immunosorbent assay (ELISA). The sera from these cattle were tested in competition enzyme immunoassays (cEIA) using two monoclonal antibodies. The results were compared to those obtained in the serum agglutination assay (SLA) and in an ELISA. The cEIAs detected more positive samples than the other tests. All sera that were positive in the SLA and in the ELISA were also positive in the cEIAs. Due to the higher sensitivity, these tests may be an alternative for the SLA.

Abstract: The occurrence of subgingival staphylococci was determined in 506 individuals with advanced adult periodontitis, 108 with early-onset periodontitis, 13 with localized juvenile periodontitis, 18 with gingivitis, and 13 with 20 failing osseointegrated titanium dental implants. Subgingival samples were collected with paper points and transported in VMGA III. The bacterial samples were plated on Staphylococcus 110 medium which was incubated in 10% CO2, and on enriched brucella blood agar, which was incubated anaerobically. Staphylococcal isolates from 94 adult periodontitis subjects were speciated using the API STAPH Trac micromethod kit system and the Bacto Staph latex agglutination test for coagulase activity. Staphylococcus epidermidis comprised 45.8% and Staphylococcus aureus 22.3% of total staphylococcal isolates. At 1 microgram/ml, in vitro resistance by staphylococci was found to tetracycline (14.4% of isolates), penicillin (4.9%), erythromycin (12.1%), and metronidazole (31.9%). Subgingival staphylococci were isolated from approximately 50% of gingivitis and periodontitis patients. No statistically significant differences were found between these patient groups in the prevalence or mean proportions of staphylococci recovered. "Periimplantitis" lesions exhibited significantly higher proportions of staphylococci (15.1%) than gingivitis (0.06%) or periodontitis (1.2%) lesions. Staphylococci may play a role in some failing osseointegrated dental implants.

Abstract: Differentiation of B lymphocytes from various peripheral lymph nodes (PLN) of mice was studied in the in vitro and in vivo conditions. Subcutaneous (SC) immunization in the foot pads with thymic independent (TI)-2 antigens such as polyvinyl pyrrolidone (PVP), R36A, and Trinitrophenyl (TNP)-Dextran failed to elicit plaque forming cell (PFC) responses from the draining popliteal and inguinal lymph nodes, whereas the splenic B cells of these animals exhibited good PFC response. Injection of unconjugated Brucella abortus (BA) along with TNP-Dextran was able to induce anti-TNP PFC formation from the draining lymph node B cells. However, PVP + BA or R36A + BA failed to stimulate such antigen specific B cell responses from the lymph nodes. Since TI antigens are metabolized slowly because of their long repeating subunits, we measured antibody responses up to 8 weeks after SC immunization with TNP-Ficoll and failed to observe any anti-TNP IgM PFC in the lymph nodes. TNP-Ficoll, in contrast to TI-1 antigen TNP-BA, did not induce IgG PFC response in the draining lymph node cells. In in vitro cultures, TNP-Ficoll induced the B cell differentiation from mesenteric lymph node (MLN) cells but not from the PLN cells. This activation of MLN B cells with TNP-Ficoll was limited to normal DBA/2J mice and was absent in the MLN cells of X-linked immune defective CBA/N mice. Finally, there was good antibody response from the inguinal and lumbar lymph node cells after an intraperitoneal immunization with TNP-BA but not with TNP-Ficoll.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: Individual milk samples and artificially constructed tank milk samples from cows with naturally occurring brucellosis were examined by the enzyme-linked immunosorbent assay (ELISA) using sonicated B. abortus S-99 antigen, and mouse monoclonal anti-bovine IgM, IgA, and IgG1 conjugates. ELISA results were compared with the results of the milk ring test using either 1 ml milk (MRT-1) or 8 ml milk (MRT-8). The ELISA using mouse monoclonal anti-bovine IgG1 conjugate was sensitive and specific. In testing individual milk samples and constructed tank milk samples containing milk with low titers in the MRT-1 the ELISA was superior to the MRT-1, and MRT-8. In testing other milk samples, the ELISA was as sensitive or slightly less sensitive than the MRT-8. From a total of 5,910 milk samples collected from cows free from brucellosis, only 24 (0.4%) samples tested positive in the ELISA. All 500 tank milk samples collected from farms negative for brucellosis tested negative in the ELISA. We concluded that the ELISA is a good substitute for the MRT-1 to detect antibodies against Brucella in milk from individual cows. When tank milk is tested for antibodies against Brucella, however, both the MRT-8 and the ELISA should be used.

Abstract: During an investigation of bovine brucellosis in Iran, conducted by the Razi Institute over a twelve-month period, samples of serum and milk were collected simultaneously from 6,472 cows in eight infected herds for serological and bacteriological testing. A total of 1,056 cows were serologically positive and 1,632 of 6,472 milk samples were positive to the milk ring test (MRT). Culture of the positive milk samples yielded 397 isolates of Brucella, 119 of which came from the 5,686 seronegative cows. The isolates belonged to Brucella abortus biotypes 2 (one isolate), 3 (356 isolates) and 9 (40 isolates).

Abstract: Enzyme-immunoassays using lipopolysaccharide as antigen were developed for the detection of bovine IgG1, IgG2 or IgA Brucella antibodies (Ab) in milk. The results of these tests were compared with those of the milk ring test (MRT) by analyzing 3212 bulk milk samples from farms located in regions where brucellosis is prevalent. Among the 105 herds detected by ELISA and/or MRT, 29 infected herds were detected by ELISA only. The 40 MRT-positive herds were also ELISA positive. Five herds became infected during the study and were detected by ELISA 15 days to 6 months prior to detection by MRT. The ELISA IgG1 titration (IgG1 ELISA) detected 92.8% of the herds found positive in the three ELISA assays. The concomitant use of IgA ELISA raised the sensitivity to 100% but slightly decreased the specificity. The IgG2 ELISA did not improve the diagnosis. The sensitivity of MRT and IgG1 ELISA was compared by testing successive dilutions in negative milk of 110 individual MRT positive milks samples. On average, IgG1 ELISA was 22 times more sensitive than MRT.

el-Daher N, Na'was T, al-Qaderi S. The effect of the pH of various dairy products on the survival and growth of Brucella melitensis. Annals of tropical medicine and parasitology. 1990 Oct; 84(5); 523-8. [PubMed: 2256775].

Abstract: In this study we report the effect of the pH of various dairy products on the survival and growth of Brucella melitensis. The growth patterns of B. melitensis in broth media at different pHs (ranging between 3 and 9) were studied for up to four weeks, to standardize the growth of the organism at each pH. These growth patterns were compared with those of the same organism growing in different dairy products [milk, soft cheese, yoghurt, and buttermilk (shaneena)] under the same growth conditions. This showed that B. melitensis could survive for more than four weeks in broth at a pH of greater than or equal to 5.5, was inhibited in less than three weeks at pH 5 and in one day at pH 4, but could not survive in a pH of less than 4. In dairy products there was a marked drop in the total viable count, and the organism could not be detected after short periods of time. After 72 hours B. melitensis could be detected only in soft cheese, but it was not detectable in any of the dairy products tested after 96 hours. Thus, this study has shown that the survival of B. melitensis in the different dairy products was inversely proportional to the pH of the product.

Abstract: Several strains of Escherichia coli O157:H7 artificially inoculated into vegetables and dairy products were recovered on hydrophobic grid membrane filters and enumerated by an enzyme-labeled antibody assay. The mean of the recoveries from 12 fresh vegetables was 108.8%, whereas that from 10 dairy products was 93.2%. Modified tryptic soy broth at 43 degrees C with shaking at 100 rpm provided optimum recovery of the organism from meat, with a sensitivity of less than or equal to 1 CFU/g, which is 10 times more sensitive than direct plating. The method performed equally well with vegetable and dairy products. Tryptic soy broth, however, under the same conditions gave the best results for fecal samples. Of 22 asymptomatic dairy cattle, reported as having positive Brucella titers when assayed with polyclonal antibodies, eight were found to contain E. coli O157 in their feces as demonstrated by the enzyme-labeled antibody assay by using monoclonal antibodies. This finding may explain some of the false-positive Brucella tests.

Abstract: A 36-year-old Hispanic man came into the emergency department with nonspecific symptoms (headache, myalgias, low-grade temperature, and low white blood cell count) and was diagnosed with brucella meningitis. The patient said he had consumed unpasteurized goat's milk and cheese in Mexico, and had been treated 3 months previously for a febrile illness diagnosed as Malta fever (brucellosis). Cultures of both the blood and cerebrospinal fluid yielded Brucella melitensis. Blood agglutinin results for B abortus were positive at greater than 1:160. Unpasteurized milk and cheese are consumed in many countries where brucellosis is endemic. Emergency physicians are occasionally confronted with patients from developing countries with diseases that require rapid and specific diagnosis for optimal treatment.

Abstract: Liposomes containing aminoglycosides have been shown to enhance the killing of Brucella abortus and Staphylococcus aureus inside bovine phagocytic cells. This study examined the mechanism by which liposomes containing aminoglycoside enhance the intracellular killing of bacteria. Liposomes with entrapped aminoglycoside were found to significantly enhance the intraphagocytic killing of bacteria in bovine phagocytic cells (in vitro) when compared to free drug. Liposomes with entrapped aminoglycoside were also found to deliver significantly higher levels of aminoglycoside into phagocytic cells when compared to free drug (gentamicin) or free drug and liposomes without entrapped antibiotic. Antibiotic delivered to adherent phagocytic cells could be detected 3 days after treatment of the cells with liposomes containing aminoglycoside. No antibiotic could be detected in the supernatants of phagocytic cell cultures 3 days after treatment with liposomes containing antibiotic was only observed when the intraphagocytic bacteria were sensitive to the antibiotic entrapped in the liposomes. The rate of phagocytosis of S. aureus by cells treated with cationic liposomes (no entrapped antibiotic) did not differ from the rate of phagocytosis of control cells not treated with cationic liposomes. This study shows that the enhanced intraphagocytic killing of bacteria in bovine phagocytic cells occurs by direct delivery of entrapped antibiotic into the phagocytic cell by the liposome delivery vehicle and not by nonspecific enhancement of phagocytic cell function. Liposomes containing aminoglycoside appear to have no toxic effects on phagocytic cell function or viability in vitro.

Abstract: Sixty-three samples of the more solid material (sludge) separated from the effluent plants of dairy factories were examined for the presence of salmonellas and brucellas. Salmonellas were isolated from two samples (S. heidelberg. [1]; S. indiana [1]. No brucellas were isolated. None of the samples supported the growth of S. dublin. Salmonellas added to effluent sludge at a concentration of 10(6) organisms/ml. survived less than 70 days. The sludge from dairy factory effluent plants does not appear to be a source for the spread of salmonellosis or brucellosis.

Abstract: Stable plurilamellar vesicles (SPLVs) entrapping aminoglycosides were used to treat infections due to Brucella species (Brucella canis and Brucella abortus). SPLV-entrapped antibiotics effectively eliminated internalized B. canis in cultures of resident murine peritoneal macrophages and internalized B. abortus in cultures of resident guinea pig peritoneal macrophages. In vivo studies demonstrated that SPLV-entrapped aminoglycosides administered to B. canis-infected mice and B. abortus-infected guinea pigs effectively eliminated bacteria from infected organs. The dosage schedule used involved two intraperitoneal administrations of SPLV-entrapped aminoglycosides at three-day intervals. The results demonstrate the superiority of SPLV-entrapped aminoglycosides to free aminoglycosides in effecting elimination of facultative intracellular bacteria in vitro and in vivo. The use of SPLVs as a drug carrier has broad application to treatment of infections due to other organisms.

Abstract: Clinical epidemiological evaluations on brucellosis, done during the last five years in endemic areas in Lima (Peru), in 39 families with 232 members, have let us observe a high rate of symptomatic infection 118/232 (50.9%) and has also let us identify, as risk factors to contract the disease; age older than 10 years 97/190 (56.3%) versus 11/42 (26.1%); in younger than 10 years of age (p less than 0.05) and families with 5 or less members 40/54 (74.0%) versus 78/175 (44.5%) of more numerous families (p less than 0.001), and also to be exposed to a common source as a form to contract the disease. We observed the importance of an adequate treatment in early stages of the disease. We detected major clinical gravity in women 23/55 than in men 5/38 (p less than 0.01), specially in brucellar arthritis (p less than 0.05). Period of time between the appearance of index case and secondary cases was of 33.8 days, but range was of 1-115 days. The heterogeneous aspects of the disease between members of the same family, suggest that response of host, more than bacterial virulence, is so important in the clinical course of the disease.

Abstract: Twenty-nine patients presenting with symptoms and signs suggestive of brucellosis in the Gezira area, Central Sudan, were studied. The majority of the patients (76%) were found to have a combined infection of both Brucella abortus and Brucella melitensis with titres of 1/160 and above. The social habit of eating raw meat, e.g. raw liver or other offal with spices (Marrara or umfitfit) is found to be an important epidemiological factor in contracting the disease. Raw milk consumption, since it is not common, does not seem to have a major role in human infection. Clinically, there are no severe complications in this series. This might be a reflection of the small size of the group studied. The study showed good tolerance and excellent response to the combined therapy of streptomycin and doxycycline.

Abstract: Thirty cows naturally infected with Brucella abortus were treated by various routes, using free or liposomal streptomycin or a combination of liposomal streptomycin and a long-acting oxytetracycline preparation. Of 21 cows treated with liposomal streptomycin alone, 3 (14%) were culture negative and 3 had 10 or fewer bacterial colonies isolated from tissues obtained at necropsy. Thirteen (62%) cows continued to shed organisms in udder secretions and were considered treatment failures. Of 9 cows that were given a combination of liposomal streptomycin and long-acting oxytetracycline, 5 (56%) were cured, 3 had 10 or fewer colonies on culture plates of tissue after necropsy and only 1 continued to shed B abortus in udder secretions after treatment. Eleven cows were given streptomycin liposomes by intramammary infusion with or without IM administration of long-acting oxytetracycline. The most effective regimen consisted of 2 intramammary infusions of streptomycin liposomes and 2 doses of oxytetracycline administered IM. Of 5 cows treated thusly, 2 were cured and all others had fewer than 10 B abortus colonies isolated from tissues obtained at necropsy.

Abstract: This study investigated the presence of Brucella antibodies in serum and milk obtained from camels in Kuwait. Brucella strains were also isolated from the foetus using standard technique (Webridge Lab Techniques). Three serological tests for serum were adopted. These tests were Rose Bengal Plate Test (RBPT), the Serum Tube Agglutination Test (SAT) and the Complement Fixation Test (CFT). The RBPT was used for all sera samples, and both SAT and CET were used for the positive RBPT. Camels that showed a titer of 1:40 in SAT or 1:5 in CFT or greater were considered positive. Thirteen of the samples examined were found positive by CFT (at 1:5); by SAT, they showed a titer of 1:20. One serological test, the Milk Ring Test (MRT), was used for milk. Here 3 and 2 were considered positive reactors but 1+ was considered suspicious. We were unable to isolate the Brucella organism from Sedemine and Cream of milk, but we isolated them from Foetus Brucella abortus and it is confirmed by Webridge Laboratory, U.K. It is Brucella abortus (Biovar 1). The prevalence rate was 14.8% from serum by the CFT and RBPT methods and 10.8% by the SAT method. For milk, the prevalence rate was 8.0%. Two Brucella abortus were isolated from 5 foetuses.

Abstract: A dipstick suitable for immunoassay procedures has been developed, and its utility demonstrated by the development of tests for the determination of immunoglobulin levels in newborn bovine blood, and for the detection of bovine antibodies against Brucella abortus. The dipstick is of simple design, consisting of three components: a polycarbonate membrane, an adhesive, and a support material. The polycarbonate membrane is a porous filter material upon which the immunoassay is undertaken. Biomolecules bind to the membrane with sufficient affinity to permit the development of useful immunoassay procedures; however, this membrane exhibits low non-specific binding of reagents. Collectively, these properties make polycarbonate a useful material for solid-phase immunoassay procedures.

Abstract: Eight 1-year-old, goats were inoculated intravenously with Brucella abortus (B. abortus) on the day of parturition and necropsied at 28 days after inoculation. Four nursed their kids and four did not (milk was not removed from the udders). Tissues and fluids were examined by bacterial isolation, light microscopy, and serologic methods. Nonnursing goats had high titers of brucellae (less than or equal to 10(8) organisms/ml) in milk (brucellae were isolated from four of four udders), had marked enlargement of supramammary lymph nodes, and had lymphoplasmacytic and histiocytic interstitial mastitis. Immunoperoxidase staining revealed that brucellae were primarily in macrophages and neutrophils of the mammary alveolar and ductal lumens and in macrophages of the subcapsular sinuses of the supramammary lymph node. In contrast, nursing goats excreted brucellae intermittently at low concentrations (less than 10(3) organisms/ml) in milk; brucellae were isolated at necropsy from one of four udders; supramammary lymph nodes were not enlarged; and mammary lesions were not seen. Brucellae were detected in more tissues other than the udder, and serum anti-Brucella antibody titers were higher in nonnursing goats than in nursing goats. The present study indicates that the failure to nurse or release milk enhances localization and replication of B. abortus in mammary glands of goats after parturition, and that mammary gland infection may result in increased systemic spread and persistence of brucellae in the host.

Abstract: 1. EC- and National Regulations. Since 1988 the EC-regulations accept in addition to the on Agar Gel Immunodiffusion test (AGIDT) based blood serum testing of cattle herds that are filed as "free from Enzootic Bovine Leucosis" the use of ELISA for this purpose. The regular testings in dairy cattle herds can be done alternatively with single or pooled milk samples, in other herds with pooled blood sera using ELISA. General condition is only a minimal sensitivity of the test to detect the European EBL Antibody Standard ("E4") in a dilution of 1:10 in negative serum or 1:250 in negative milk. Adequate national regulations are in preparation. The present limitation of pool sizes, blood maximum 50 animals without preparation steps 20, and milk after concentration treatment 50 cows is neutralized by proceedings in development of higher sensitive ELISA tests. This limitation should be canceled. Herd bulk milk samples without size limitations are accepted to be tested with "Milk Ring Test" by EC for the regular testings in filed "Brucellosis Free Dairy Cattle Herds". The alternative use of more sensitive (and more specific) ELISA tests for this purpose including the technical conditions is in a final discussion. 2. Scientific-Technical Base for Using the Chances of the Proceeding in the EC-Regulations. The realisation of the EC accepted or final discussed ELISA based bulk milk testing to control filed "EBL- and/or Brucellosis Free Herds" depends on some basic conditions like sensitivity, specificity, and variability of the ELISA systems. Field trials of more than 20,000 bulk milk samples in case of Brucellosis and more than 2,000 in case of EBL show the feasibilities and the limits of the ELISA systems in defining the status of the herds. The Brucellosis respectively the EBL situations of the dairy cattle herds tested in this trail were well known by history and by investigation of single animal blood samples using conventional tests. Special test run variations of pretested assays demonstrated the possibilities to define the EBL status of dairy cattle herds up to 50 lactating cows without preparation of the bulk milk sample and up 100 after concentration of the antibodies by the rennet-ammonium sulfate method. The concentration limit for detection of Brucellosis antibodies is 100 lactating cows. The bulk milk of smaller herds can be tested without concentration. On principle the evaluation of the test values bases on defined relations to a "weak positive" reference.(ABSTRACT TRUNCATED AT 400 WORDS)

Abstract: A 9-year-old Inuit boy with brucellosis is presented. The most likely source of his infection was contaminated caribou meat. Brucella suis is enzootic in Canadian caribou herds and this case indicates that natives of the Arctic are a risk group for acquiring brucellosis through the ingestion of raw caribou meat.

Abstract: We described a cloned dendritic cell, clone Den-1, that is a potent accessory cell for some B cell responses. Clone Den-1 produces a novel lymphokine that is distinct from previously described factors produced by T cells. In the present study, we compare the role of nonspecific helper factors produced by Den-1 (Den-1 SN) or the T cell thymoma EL4 (EL4-SN) in promotion of B cell plaque-forming cell (PFC) responses to a variety of antigens. We find that the antigen in culture determines the B cell requirement for dendritic and/or T cell factors. B cell PFC responses to TNP-Brucella abortus (BA) and TNP-lipopolysaccharide (LPS) are greatly increased by EL4-SN but show little, if any, enhancement with Den-1 SN. Responses to TNP-polyacrylamide are reconstituted by either Den-1 SN or EL4-SN, whereas responses to TNP-Ficoll, TNP-dextran and TNP-levan are reconstituted by Den-1 SN and are much less sensitive to factors present in EL4-SN. Responses to SRBC require the presence of both Den-1 SN and EL4-SN. We also show that the time at which Den-1 SN must be provided to the B cell is dependent on the antigen in culture. Our findings are discussed in terms of present classification of antigens based on their ability to stimulate various B cell subpopulations.

Abstract: A 21-year-old butcher presented with fever and severe thrombocytopenic purpura, and was found to have acute brucellosis. After treatment with tetracycline, gentamicin, and prednisone, the thrombocytopenia resolved over 14 days. A brief review of thrombocytopenic purpura associated with brucellosis is presented.

Abstract: Disease control must be part of the management system in any beef cattle enterprise where improved net returns are a constant goal. A current view of the importance of disease is reflected in the research portfolio of the Australian Meat and Live-stock Research and Development Corporation (AMLRDC), where only 14% of projects relate to cattle health problems. Our relative freedom from epidemics is partly responsible for this balance, but the profession should continue to be alert to the range of actual or potential threats to the national herds and those predisposing factors, both managerial and environmental, that influence their effects. In Australia with its crucial beef export component, several priority areas can be identified. They are (i) completion of the Brucellosis and Tuberculosis Eradication Campaign (BTEC) (ii) improved knowledge of the effects of production of subclinical disease including deficiency states (iii) better understanding of the infertility complex from conception to birth. Congenital viral infections such as mucosal disease and the arbovirus group are slowly emerging from their epidemiological obscurity (iv) constant awareness of the potential threat of exotic diseases and defensive methods available to meet them. There is a particular need to understand the epidemiology of the large tropical cattle herd, which interfaces with South-East Asia. Our comparative freedom from epidemics should not cause any relaxation of vigilance by the veterinary services in the private or public sectors. Nor should government be allowed to minimise the critical role of the veterinary profession in our defense against animal disease (v) increased liaison with our trading partners in livestock exports.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: Brucellosis, a bacterial infection, is rarely seen in the United States. It occurs mostly in people who work with domestic animals and animal products. However, this patient became infected while in Yugoslavia, and diagnosis was delayed by the gap between appearance of symptoms and positive results of serologic tests. Brucellosis should be suspected in a patient with unexplained fever, especially if he or she has traveled to a country where unpasteurized dairy products are common.

Fricker CR, Park RW. A two-year study of the distribution of 'thermophilic' campylobacters in human, environmental and food samples from the Reading area with particular reference to toxin production and heat-stable serotype. The Journal of applied bacteriology. 1989 Jun; 66(6); 477-90. [PubMed: 2753842].

Abstract: The incidence of 'thermophilic' campylobacters in foods and environmental samples has been studied over a two-year period. Of 781 environmental samples, 529 (67%) were found to contain campylobacters, and campylobacters were isolated from 835 (39%) of 2116 food samples. Sewage was almost always contaminated with campylobacters (96.6% of samples) and of the food samples both poultry (55.5%) and offal (47.0%) were commonly contaminated. Determination of the heat-stable serotypes of all strains isolated from these sources and of 921 strains isolated from human faeces showed that there was a wide distribution of serotypes in most types of sample. Serotype Pen 2 was the commonest type found in human faeces (18.9%) and it was also commonest in offal (21.3%), beef (40.0%), sewage (17.7%) and was the third commonest type in poultry. A comparison of culture media and conditions for optimal production of both cytotoxic and cytotonic enterotoxins showed that Brucella Broth incubated under microaerobic conditions for 24 h at 42 degrees C was suitable for both toxins. Detection of cytotoxic activity was most sensitive using HeLa cells. The sensitivities of two ELISA systems and a Chinese Hamster Ovary tissue culture assay for detection of cytotonic enterotoxin were comparable. Not all strains isolated from cases of enteritis in human beings produced toxin; 23.1% produced cytotonic enterotoxin and 17.5% produced cytotoxin. There was no correlation between serotype and toxin production. The wide distribution of campylobacters, indistinguishable from those isolated from cases of enteritis in human beings, leads us to conclude that simplistic statements suggesting that one particular type of food is primarily responsible for cases of human disease should not be made.

Abstract: From 1977-1986, a total of 331 laboratory confirmed cases of human brucellosis were reported in Texas. The annual number of cases ranged from 13 in 1977 to 84 in 1983. Males accounted for 66% of the cases. Cases ranged in age from one to 92 years, with 57% between the ages of 20 and 49 years. Brucella melitensis infections accounted for 66% of the bacteriologically confirmed cases. Infection with Brucella canis was identified in four patients. Annual incidence rates were higher in Hispanics in each age and sex group when compared with whites and blacks. Epidemiologic features of cases reported during 1977-1981 are dissimilar to features of cases reported during 1982-1986. In the first five years, a majority of cases (82%) were males, and a majority (54%) were white. Exposure to cattle or swine was reported for 72% of the cases. In the second five years, a majority (72%) of cases were Hispanic, and only 55% were males. Ingestion of unpasteurized goat milk products was reported for 67% of the cases during 1982-1986.

Abstract: Liposomes with entrapped gentamicin were used to treat mice with infection attributable to Brucella melitensis. Liposomes bearing positive charge and formed by egg yolk lecithin, cholesterol, and stearylamine were effective in the elimination of B melitensis residing in liver and spleen. Negatively charged liposomes, formed by egg yolk lecithin, cholesterol, and dicetyl phosphate were also effective in suppression of the infection in liver, but were less so in suppression of the infection in the spleen. Free gentamicin was less effective than the encapsulated antibiotic. At 20 hours after administration of gentamicin encapsulated in liposomes, the gentamicin concentrations in liver and spleen were similar, regardless of the charge of the liposomes--neutral, positive, or negative. However, positively charged liposomes were more efficient than were other liposome types for the treatment of brucellosis caused by B melitensis.

Abstract: Brucellosis involving the nervous system usually shows meningomyelitis and/or radiculoneuritis and can by their clinical appearance not be differentiated from other chronic proliferative diseases of the nervous system. Sporadic cases can only be suspected on clinical grounds if a previous exposition is known. The cerebrospinal fluid showing a proliferative or granulomatous cytological picture is strongly suggestive of the diagnosis, which is confirmed by two rising titer values in the complement binding reaction for Brucella-specific antigen. The agglutination method of Widal is not reliable due to blocking incomplete antibodies. A 46 year-old man developed a transverse myelitis within several months accompanied by fluctuating meningeal signs, segmental irritation and transient cranial nerve palsies. No involvement of other organs and no general symptoms of infectious diseases were seen throughout the clinical course. Most importantly, starting treatment as early as possible is decisive for the outcome using a combination of streptomycin, sulfonamides, gentamycin, rifampicin and tetracyclines.

Abstract: Bacterial isolates, originating from 36 subclinically infected quarter milk samples, were labelled with 75Se and checked for cream-rising at various temperatures in a system analogous to the ABR test ("Abortus Bang Ringprobe"; the cream-rising test based on stained brucella organisms for detection of brucellosis). Diagnostic specificity and sensitivity were analyzed in experiments where labelled bacterial isolates were mixed with a number of quarter milk samples with known bacteriological status as well as samples from healthy control quarters. Creaming at 37 degrees C resulted in specific "recognization" as the bacterial isolates showed preferential flotation in the milk samples from which they had been isolated as well as is milk samples harbouring the same bacterial species. At lower creaming temperatures, the specificity was lost since all the isolates became concentrated in the cream phase irrespective of the milk sample. When comparing the specific recognization by cream of the respective bacteria, bacterial species vary: The prospects for developing diagnostic cream-rising tests for Streptococcus agalactiae, Staphylococcus aureus and Escherichia coli seems promising, but less so for coagulase-negative staphylococci, Streptococcus dysgalactiae, and Streptococcus uberis. The mechanism behind the cream-rising of labelled bacteria at 37 degrees C seems to lie in specific fat globule membrane-bound immunity of IgA type. Therefore the milk fat globules from chronically infected quarters function as absorbents for the respective isolates. Flotation of bacteria with cream indicates an in vivo mechanism enabling bacteria to invade the upper parts of milk ducts within the udder.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: Sodium polyanetholesulfonate (SPS) is used as a routine supplement to blood culture media to enhance recovery of microorganisms, but it inhibits the growth of Peptostreptococcus anaerobius, Neisseria meningitidis, Neisseria gonorrhoeae, and Streptobacillus moniliformis. Comparative clinical blood culture studies at the University of Colorado Hospital suggested that SPS also inhibits the growth of Gardnerella vaginalis. We inoculated 16 blood culture isolates of G. vaginalis into 11 blood culture media containing SPS or sodium amylosulfate, with and without gelatin. In the absence of gelatin, only brain heart infusion and thiol broths with SPS supported the growth of more than five strains of G. vaginalis, whereas all media except Bactec 6B and 7C and brucella broths recovered most isolates with SPS and gelatin or with sodium amylosulfate alone. We conclude that SPS inhibits the growth of G. vaginalis in blood culture media but that this inhibition is medium dependent and can be overcome by supplementation of most media with gelatin.

Abstract: Milk immunoglobulins were extracted from the stained cream layer of positive milk ring tests from experimentally inoculated or naturally infected cows. IgA was always found, associated with IgM in most cases (15/17) and with IgG in a smaller number of cases (11/17). An additional incubation at 20 degrees C for 18 h gave clearer positive and negative results and a lower limit of detection than that of the usual milk ring test.

Abstract: Pregnant goats in midgestation (7 to 16 weeks) were conjunctivally exposed to Brucella abortus strain 2308 to evaluate their applicability as an animal model for bovine brucellosis. Brucellae were isolated from uterine fluid and/or placental specimens of 10 of 12 does at parturition. Six of the 10 infected does delivered dead fetuses and 1 of the 10 delivered live, premature twins. Dead fetuses typically contained brucellae in multiple tissues, whereas brucellae generally were not isolated at birth from live kids. After parturition, B abortus was excreted in the milk and uterine fluids of the infected does. At necropsy (6 weeks after parturition), organisms in the doe were primarily in the uterus and in the lymph nodes that drained the mammary glands, uterus, and head. Brucella abortus was most often isolated from the cranial lymph nodes of neonates that had remained with their dam for 6 weeks after parturition. Serum anti-Brucella antibody concentrations were determined by use of standard tube agglutination, mercaptoethanol agglutination, Rivanol plate tests, card tests, complement fixation, hemolysis-in-gel tests, and an enzyme-linked immunosorbent assay. Serologic responses were detected 2 to 3 weeks after exposure and remained detectable until parturition. Antibody titers increased after parturition in does shedding B abortus at parturition. Anti-Brucella antibody was not detected in neonates before colostrum intake. The neonate's postcolostral titers were similar to those in the dam at the time of parturition. Milk anti-Brucella antibody was detected in milk (milk ring test) from infected and noninfected mammary glands.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: Twenty dairy heifers each consumed 350 mg of chlortetracycline/day in their feed. Four tests were performed on serum specimens from these and 20 control calves after vaccination with Brucella abortus strain 19. The numbers of positive test results on the card test and mean titers on the tube and rivanol agglutination and complement-fixation tests were compared in the 2 groups. Using the rivanol and complement-fixation tests, there were differences in the mean titers at weeks 5 and 6 after vaccination, but by week 10, differences were not found. The results suggest that addition of low concentration of chlortetracycline in feeds have minimal effects on postvaccinal serologic reactions determined after strain-19 inoculation.

Abstract: In four separate trials, 10 coyotes (Canis latrans) which had been individually fed mascerated bovine placental tissue experimentally inoculated with Brucella abortus strain 2308 were placed in a 1 ha isolation area with six parturient, non-B. abortus vaccinated, Brucella spp. seronegative Hereford heifers. During the second trial, three of the heifers became Brucella spp. seroreactive (as determined by the card, standard agglutination tube, rivanol, complement fixation, and enzyme labeled immunosorbent assay tests) 14 days after exposure to the B. abortus infected coyotes. Five of 10 coyotes in the second trial seroconverted to Brucella spp. positive by day 14 postexposure (PE) and by day 27 PE seven of the 10 coyotes were Brucella spp. reactive. The three Brucella spp. seroreactive heifers subsequently aborted on days 35, 64 and 66 PE and B. abortus strain 2308 was isolated from vaginal swabs, milk, and placental and fetal tissues. There was no serologic or bacteriologic evidence of transmission from B. abortus infected coyotes to the susceptible cattle recorded in the other three trials.

Abstract: Isolation and identification of Brucella antigenic determinants important to cellular responses have been difficult. In this study, bovine peripheral blood mononuclear (PBM) cells from cattle vaccinated with Brucella abortus 19 proliferated to extracted bacterial proteins blotted onto nitrocellulose. Proteins were extracted from gamma-irradiated B. abortus 19 with a sodium dodecyl sulfate extraction buffer. The extracted proteins were separated electrophoretically by sodium dodecyl sulfate-polyacrylamide gel electrophoresis prior to electroblotting onto nitrocellulose. Nitrocellulose sections corresponding to individual lanes of the gel (containing all separated proteins) were then cultured with the PBM cells. Primary and secondary stimulation responses of the PBM cells with the whole protein blots were similar kinetically to the responses of the PBM cells stimulated with whole irradiated B. abortus 19 or with whole irradiated B. abortus 19 blotted onto nitrocellulose. Although lipopolysaccharide was determined to be associated with the extracted proteins and transferred onto the blots, the lipopolysaccharide did not stimulate cellular proliferation, as indicated by the antigen-specific secondary responses. Stimulating PBM cells with portions of the blot containing high (greater than 45,000)-, medium (25,000 to 45,000)- or low (25,000)-molecular-weight proteins demonstrated that the responding cells were specific only to the proteins of corresponding molecular weights. These results indicate that cellular responses to individual proteins can be studied without cloning the bacterial genes or purifying the individual proteins.

Abstract: Brucella abortus strains were isolated from bovine tissue and milk samples from seven Ontario herds. The isolates were characterized by colonial morphology, requirement of CO2 for growth, lysis by Tbilisi phage, biochemical tests and agglutination in monospecific sera. They resembled B. abortus biotype 2 (on the basis of sensitivity to thionin and basic fuchsin) and biotype 4 (on the basis of agglutination with anti-Brucella "M" but not anti-Brucella "A" absorbed sera). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of these isolates and B. abortus biotypes 1, 2 and 4 showed similar profiles. Immunoblots with anti-A and anti-M absorbed sera showed different antigenic regions reacting with the specific sera and also confirmed that the atypical B. abortus isolates were serologically similar to biotype 4.

Abstract: A sanitary and technological diagnosis of the goat cheese rural process was carried out. The purpose was to obtain more information for the planning of a program aimed to the improvement of this small agroindustry. Samples of milk, curdle, dry abomasum, rennet, water and cheese of 10% of the small industries of two rural villages in two agricultural seasons, were taken. Moreover, dilutions of the utensils and goat udders were prepared. The samples were subjected to microbiological analysis of mesophilic aerobic bacteria count, most probable number of total and fecal coliforms, and detection of Staphylococcus aureus coagulase (+), Salmonella typhi and Brucella melitensis. Proximate chemical analysis and determinations of sodium chloride and titratible acidity in milk, cheese, dry abomasum and rennet, were carried out. Goat milk was also subjected to analysis of density. It was found that significant sanitary failures are present during th whole goat cheese process, although the highest bacteria contamination occurred at the milking, curdling and filling stages. These are characterized by excessive handling and absolute lack of hygiene. The pathogen B. melitensis was absent; therefore the causes of poisoning were attributed to the toxin produced by S. aureus and to the significant count of fecal coliforms found in the goat cheese. Even though the goats are fed under a poor feeding system, the milk presented a normal physical and chemical composition. Nevertheless, protein and fat matter losses occur during cheese preparation, as a result of handling practices and lack of process control.

Abstract: Complement fixing (C.F.) antibodies to Campylobacter jejuni were detected in 83 slaughterhouse workers and 83 blood donors. Workers were aged 18-65 years (mean, 41.7 +/- 12.3) and had worked in the slaughterhouse for 2-40 years (mean, 17.5 +/- 5.1). C.F. antibodies were detected according to Mosimann's method and by including five antigens: Campylobacter jejuni, Yersinia enterocolitica types 03 and 09, Yersinia pseudotuberculosis and Brucella. Positive titers were found in 12.1% of workers and in 2.4% of control subjects (p less than 0.01); values ranged from 1:10 to 1:40. Frequent and close contact with animals or their products was significantly associated with seropositivity. No association was found with the time of employment. Sixty per cent of seropositive workers referred rheumatological symptoms. These findings confirm that slaughterhouse workers exposed to potential sources of C. jejuni have elevated titers of antibodies. Attention has, therefore, to be focused on breaking the chain of transmission as a means of control.

Abstract: We report a case of a distinctive dermatitis in a dairy worker exposed to Brucella abortus while manually delivering an infected calf. The rapid onset of pruritus, the strict limitation of lesions to the upper extremities, and the negative results on growth of culture for microorganisms make direct bacterial infection an untenable cause in this case. Allergic hypersensitivity is the probable mechanism, manifesting itself initially as contact urticaria. Coincidentally our patient also had systemic brucellosis, the symptoms of which antedated and were unrelated to his cutaneous disease.

Abstract: A 45-years-old Greek patient developed septicaemia in his 10th year of hemodialysis treatment. Clinical investigation was directed first on bacterial infection of the arteriovenous shunt, on urosepsis or bronchopulmonary infection. Then, serologically (Widal's reaction and ELISA-test) and in different blood-cultures Brucella melitensis was detected as causative agent for Malta fever (Febris undulans). The focus of infection is suspected in unpasteurized sheep-milk cheese of mediterranean origine, wherein Brucella species can survive for months. The patient was treated effectively by a combination of amoxycilline and clavulanic acid.

Abstract: The clinical and pathological findings in a patient who had acute hepatitis caused by Brucella melitensis are described. Antibiotic therapy induced a good clinical and biochemical response, although a relapse occurred. Brucellosis must be considered in the differential diagnosis of pyrexia of unknown origin, particularly if associated hepatic involvement is present. A careful occupational and travel history is essential.

Abstract: The brucella antibodies were investigated in 132 different milk samples and 150 human sera by the Ring Test. The guinea-pig tests were made to determine the antibody level and microorganism isolation using one seropositive milk sample. Increase of the antibody titre was observed. Brucella germs from blood, spleen and liver of guinea-pig didn't multiply.

Abstract: Bulk tank milk samples which were collected twice with an interval of 2 months from 2103 herds were tested for brucellosis by employing the brucella ring test. Farmers involved were all supplying industrial milk to the National Co-operative Dairies. A questionnaire was circulated to these farmers in which they were asked to indicate how heifer calf vaccination with strain 19 vaccine was practised on their properties. Of the herds tested, 18.1% could be regarded as infected. This figure varied from 8.9% in the Potchefstroom area to 30.9% in the northern districts of the Hoopstad area. An important cause of this rate of infection should be sought in improper calfhood vaccination. At least 9% of the respondents did not practice heifer calf vaccination whilst another 8.1% only commenced with vaccination during 1985 or later. About 12.4% of farmers commenced heifer calf vaccination prior to 1970.

Abstract: Spleen cells from male (CBA/N x DBA/2) F1 hybrid mice do not significantly respond to in vitro stimulation by trinitrophenyl-conjugated polyacrylamide beads (TNP-PAA), whereas the same antigen elicits high PFC responses in female F1 hybrid cells. Therefore, this antigen could be classified as a T-independent type 2 (TI-2) antigen. When male spleen cells were co-stimulated by TNP-PAA and TI type 1 antigen, either LPS or Brucella abortus, they produced vigorous anti-TNP responses. A similar increase of the in vitro responsiveness of male F1 hybrid spleen cells to TNP-PAA antigen was provoked by the addition of supernatants from P 388-D1 cells stimulated by muramyl-dipeptide (MDP) mainly containing interleukin-1 (IL-1) or supernatants from phorbol 12-myristate 13-acetate (PMA)-stimulated EL-4 cells that contained T-cell factors. The PFC response to another TI-2 antigen, TNP-Ficoll, was also significantly enhanced after co-stimulation by P 388-D1 supernatants. The response to TI-2 antigens being macrophage dependent, the influence of supernatants of peritoneal macrophages from male and female F1 hybrids incubated with TNP-Ficoll on the PFC response of normal DBA/2 mouse spleen cells to sheep erythrocytes was assessed. It was found that macrophage supernatants from female hybrids regularly increased by more than two times this anti-SRBC PFC response, whereas macrophage supernatants from male F1 hybrids did not. Moreover, in a specific proliferation test measuring IL-1 activity, when macrophage supernatants from female F1 produced a 13-fold increase of thymidine incorporation, supernatants from male F1 only produced a three-fold increase. It is concluded that, in addition to the known defects of B cells from Xid mice, their macrophages are also defective.

Abstract: Single serum samples from 559 volunteers from a Texas Gulf Coast area were examined for vibriocidal antibody to Vibrio cholerae O1 (biotype El Tor, serotype Inaba) by a microtiter method. Elevated levels of vibriocidal antibody were present in 14% of the subjects. Also, 6.8% of the subjects had elevated levels of antibody to the enterotoxin of V. cholerae O1 by the immunoglobulin G enzyme-linked immunosorbent assay. Recent infection, defined on the basis of elevations in both vibriocidal and antitoxin antibodies, had occurred in 1.3% of the subjects. When subjects who reported Brucella infection, travel to a cholera-endemic area, and/or cholera vaccination within a year of the study were removed from the analysis, a prevalence of recent infection of 0.89% was obtained. Significantly higher titers of vibriocidal antibody were found in those with exposure to seawater (fishermen, shrimpers, merchant marines, and dock workers) than in those without such exposure (P less than 0.005). Furthermore, titers of antitoxin antibody were significantly higher in those who consumed shellfish than in nonconsumers. Finally, titers of vibriocidal antibody were significantly higher in Vietnamese subjects than in non-Vietnamese subjects. The results of this study indicate that an endemic focus of infection with V. cholerae occurs in this area.

Abstract: A survey of the most common food borne infections (salmonellosis, tuberculosis, brucellosis, campylobacteriosis) is followed by a description of the most common food borne intoxications (botulism, staphylococcal intoxications, biogenic amines, toxic algae in mussels, i.e. "Paralytic Shellfish Poisoning").

Abstract: The susceptibilities of 36 isolates of the Bacteroides fragilis group to ceftizoxime, cefoxitin, clindamycin, and metronidazole were determined by using the National Committee for Clinical Laboratory Standards agar dilution reference method and a broth microdilution method using anaerobe, brucella, Schaedler, and brain heart infusion broths. MICs that were greater than or equal to fourfold higher or lower than those of the reference method were considered significant. Major and minor discrepancies in susceptibility interpretation (SI) were also noted. Ceftizoxime showed the greatest number of variations and SI discrepancies. In 72% of the cases, MICs in broth were greater than or equal to fourfold lower than those obtained by the reference method, resulting in 33% of the major and 22% of the minor discrepancies in SI. A total of 53% of the isolates were resistant to ceftizoxime by the reference method, but only 11 to 17% were resistant in the various broths. Significant variations in MICs of cefoxitin occurred in 19 to 22% of the isolates; 17 to 19% of the isolates showed major discrepancies and 31 to 58% showed minor discrepancies in SI. A total of 58% of the isolates were resistant to cefoxitin by the reference method, but only 19 to 28% were resistant in the various broths. Significant variations with clindamycin in broths ranged from 32 to 53% and resulted in 3 to 8% of the isolates showing major discrepancies and 33 to 44% showing minor discrepancies in SI. Metronidazole yielded significant variations in MICs in 6 to 28% of the isolates, but no major or minor SI discrepancies were noted. This study indicates that significant differences in susceptibility results, which appear to be method related, can result when isolates of the B. fragilis group are tested. Therefore, studies correlating in vitro results, determined by various methods, to clinical outcome are essential.

Abstract: The present study was undertaken to compare the effects of two adjuvants, SGP (a starch-acrylamide polymer) and Quil A (purified saponin), with that of aluminum hydroxide (Al(OH)3) on murine primary antibody responses to T-independent (TI) and T-dependent (TD) antigens. All three adjuvants augmented the responses to the TD antigens, dinitrophenyl-keyhole limpet hemocyanin (DNP-KLH), and sheep erythrocytes (SRBC). SGP was the most potent adjuvant and increased the primary IgG response to DNP-KLH as much as 90-fold. Quil A and Al(OH)3 had comparable effects on the primary response to DNP-KLH, but Quil A was less effective than Al(OH)3 for augmenting the primary response to SRBC. Quil A and SGP both augmented the primary IgM and IgG responses to trinitrophenyl-lipopolysaccharide (TNP-LPS), TNP-Brucella (TI-1 antigens), and TNP-Ficoll (TI-2 antigens). Al(OH)3, like most commonly used adjuvants, had little or no effect on responses to TI antigens. The kinetics of the response to TNP-Ficoll was altered by SGP, since peak responses were maintained for at least 7 days, while the response to TNP-Ficoll alone peaked on Day 4 and had declined considerably by Day 7. Both SGP and Quil A could augment responses to both optimal and suboptimal doses of antigen. The adjuvant activity of SGP was diminished, but still effective, when smaller amounts of SGP were used with the immunizing antigen, and all three adjuvants were able to augment primary responses when given in separate injections from the antigen. These results demonstrate that SGP is a very effective adjuvant, and show that both Quil A and SGP have a unique ability to increase antibody responses to TI antigens, suggesting that their effects may be mediated at least partially through B cells.

Abstract: The backgrounds to and possibilities of serological studies for the purpose of establishing are reviewed, all this with regard to bovine disease. The serology of the virus infections infectious bovine rhinotracheitis (IBR), bovine viral diarrhoea (BVD), para-influenza virus 3 infection (PI3), bovine respiratory syncytial virus (BRS) and Chlamydia is discussed in greater detail. Likewise, the interpretation of serological findings in cases of brucellosis, leptospirosis and Johne's disease is dealt with. In addition, attention is paid to the results of serological testing in cases of lung-worm disease and fascioliasis.

Abstract: 156 Milk samples were obtained from farms, dairies and streets and brucella antibodies were searched by serologic methods. Results were compared by whey antiglobulin test (WAGT) findings. In order to established the Brucella antibodies presence according to WAGT at the performed serologic tests, milk Ring test (MRT) was found the most sensitive and pratic test. The application of milk and serum (whey) to slide aglutination tests were not seen reliable.

Abstract: A total of 2,698 dairy herds were surveyed in 1981-1982 in New South Wales and north eastern Victoria in a review of the methods used to monitor them for the presence of Brucella abortus. The methods used to monitor dairy herds were testing of all breeding cows over 1 year of age using the rose bengal test (RBT) and complement fixation test (CFT), the bulk milk ring test (BMRT), and testing of blood samples collected at abattoirs using the RBT and CFT. The surveyed herds had at least one whole herd test, and BMRT was done at regular intervals in the period of the survey. Of the 99 (3.7%) herds that reacted to the BMRT, 91 (3.4%) herds had false positive reactions and 8 (0.3%) herds were declared infected on follow-up herd testing. False-positive reactions were obtained in 22 herds on more than one occasion. Common causes of false positive reactions to the BMRT were thought to be previous vaccination with Strain 19 and sampling in very early or late lactation. Of the 98 (3.63%) herds that reacted to the whole herd serological tests, 80 (2.96%) herds had false-positive reactions and 18 (0.67%) herds were declared infected. Strain 19 vaccination was thought to be an important cause of false-positive reactions. Fifty-three (2.0%) herds showed suspicious reactions on abattoir monitoring but none was declared infected on follow-up testing. Of the 18 herds with infected or equivocal status, the BMRT identified?(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: A serological survey was carried out in Bénin in order to assess the rate of brucellosis infection among exposed workers (workers in slaughtering-houses and breeders). 221 sera were tested with rose Bengale test, Wright sero-agglutination test, indirect immunofluorescence test and counter-immuno electrophoresis (brucelline). The percentage of positive sera among exposed workers is 17,7%. The rose Bengale and immunofluorescence tests combination permits complete detection of positive sera. These results suggest the existence of human brucellosis in Bénin and shows the necessity of a national control programme adapted to the socio-economic problems of this country.

Abstract: Blood samples obtained from 256 workers of an abattoir were tested for brucellosis antibodies and 11.7% were found reactive. The highest rate was 44.2% among those who had contact with blood and viscera, and 26.1% among workers at slaughter section. Four tests were used: plaque agglutination (PA), tube agglutination (TA), card test (CT) and complement fixation (CF). The highest number of positive reactions was obtained by the PA and TA tests, with a complete coincidence in both results. When both test titers were the same or less than 50 IU, CF and CT yielded less percentage of positive results. When titers were higher than 50 IU, CT but CF showed concordance between the results.

Abstract: To begin with, a brief historical review of a number of features of bovine fertility is presented in the present paper which is part of the series 'Papers of Yesterday and Today'. This is followed by a discussion of the current situation. All this is done on the basis of a paper by Stockfleth which appeared in 1878. As a result of the eradication of venereal infections and brucellosis, becoming pregnant as well as pregnancy run a course associated with less problems than it was one hundred years earlier. Errors of herd management are immediately reflected in fertility. When animals fail to stay pregnant (abortion), a large number of possible causes may be suggested. Infections are among the most important of these causes. Finally, attempts are made to look into the future. Becoming pregnant will probably run a more difficult course in view of the fact that getting the cycles going post-partum in highly productive cows gives rise to additional problems. As regards staying pregnant, considerable changes are not anticipated.

Abstract: Selected aspects pertaining to the cause and pathogenesis of diskospondylitis were investigated in a kennel of 45 Airedale Terriers. Diskospondylitis was detected via spinal radiography in 17 male and 14 female dogs. Bacteria isolated from the coat, vagina, and urine of affected Airedales were not statistically different from those isolated from clinically normal Airedales. Serologic evidence of Brucella canis infection was not detected. There was no difference in response to thyrotropin stimulation tests between affected and clinically normal dogs. Necropsy findings in affected dogs included hypercellular renal glomeruli and pulmonary arterial thrombi.

Abstract: Researchers conducted an epidemiologic investigation of an outbreak of brucellosis at a meat-packing plant, where work was interrupted by a strike. The investigation revealed that the risk of infection with brucellosis may coincide with the rate at which reactor cattle are slaughtered, a factor often overlooked in other reports which investigated outbreaks of brucellosis. The slaughter of 20 to 25 reactor cattle per day was the estimate of the critical number of reactor cattle when the risk of infection is most likely to occur. Of the 193 workers studied, 17 (8.8%) were seropositive, but only eight (4.1%) were presumptive cases. Young, newly employed workers who did not wear glasses were at greatest risk of infection.

Abstract: Australia, a major exporter of meat, has met and continues to meet the import requirements of various countries. It is free of many epizootic and zoonotic diseases and is eradicating bovine brucellosis and tuberculosis. Frequently, individual country requirements have not been relevant to the animal and public health status in Australia. Such a situation is unscientific, wasteful and unnecessary, and may divert priorities away from areas of major public and animal health significance which should be the main concern of consumers both in Australia and overseas. In recent years the Inspection Service of the Department of Primary Industry has reviewed meat inspection procedures necessary to protect public and animal health in Australia and in countries importing Australian meat. Priorities include attention to national or regional occurrence of disease in Australia and to the use of scientific principles in inspection, including disease identification, concepts of pathogenesis and effectiveness in removal of diseased tissue from meat. Revised post-mortem procedures for cattle, pigs, sheep and goats more relevant to disease occurrence and consideration of public health are described. In particular, this involves a reduction in the number of lymph node incisions. Future directions for meat inspection in Australia are postulated. Concern is expressed that the requirements of some importing countries are fixed in their legislation and consequently might inhibit desirable developments. In this context it is important that scientifically sound national codes for meat inspection of the exporting country be accepted by importing countries as providing public and animal health safeguards.

Abstract: In 1983 an outbreak of human brucellosis caused by Brucella melitensis occurred among residents of a predominantly Hispanic neighborhood in Houston, TX. The source of the infections was traced to unpasteurized goats' milk cheese imported from Mexico. Nineteen of the 31 patients who contracted the disease were members of 5 families, which included 11 children ranging in age from 2 to 17 years. Infection was generally associated with protein complaints and a paucity of physical findings. Both immunoglobulin M and immunoglobulin G agglutinins were present in acute phase sera in the majority of patients, and low titers of immunoglobulin M antibodies remained in the sera of 43% who were retested 2 years later. Trimethoprim-sulfamethoxozole was highly effective in the treatment of children with brucellosis and was associated with no relapses.

Abstract: The clinical features of acute Brucella arthritis were reported in a prospective study of 58 patients (29 male, 29 female). Polyarthritis occurred in 33, was migratory in ten, additive in 23 and symmetrical in ten. Monoarthritis of knee or hip joints occurred in ten patients, whilst the spine alone was involved in 15. Hip and spinal involvement was more destructive than peripheral joint involvement. Hotness, redness and joint effusions were not prominent features. Tissue-typing did not show any association with Brucella arthritis. Treatment with various antimicrobials was followed by recovery, but the relapse rate was lowest with combined tetracycline and streptomycin. Brucella arthritis is frequently seen in Iraq, and dairy products prepared locally from unpasteurized milk should be controlled by local health authorities.

Abstract: In a 12-month survey (June 1982-May 1983) 41 laboratories examined 2493 samples of goat's milk for colony counts and the presence of pathogens. The statutory tests for cow's milk were also applied. Surface counts of less than 10(5) organisms per ml of raw milk were given by 79% of samples at 37 degrees C and by 76% at 22 degrees C. There were less than 100 coliforms per ml in 71% of samples, less than 10 Escherichia coli per ml in 91%. Staphylococcus aureus was not detected in countable numbers in 96% of samples. Only one isolation of campylobacter was made and two of Yersinia enterocolitica. Salmonella was not detected in 2462 samples. The methylene blue test was carried out on 2368 samples and 86.7% were deemed satisfactory. No sample was Brucella ring-test-positive. Experiments on the survival and growth of six food poisoning organisms in stored goat's milk showed that Bacillus cereus, Staph. aureus, Salmonella typhimurium and Y. enterocolitica survived quite well and multiplied at the higher storage temperature of 30 degrees C. Clostridium perfringens only increased 10- to 100-fold while Campylobacter jejuni did not grow. The results of the survey indicate that any problems with goat's milk relate to poor hygiene during production rather than transmission of organisms from the goat herself.

Abstract: In vitro intraphagocytic killing of Brucella abortus in bovine mononuclear leukocytes was enhanced by cationic, anionic, and neutral multilamellar liposomes-containing gentamicin. Free gentamicin not entrapped in liposomes. and liposomes without antibiotic did not enhance intraphagocytic killing of B. abortus in bovine phagocytes. In vivo killing of B. abortus in guinea pigs was also enhanced by liposomes-containing gentamicin when compared to free gentamicin. Liposomes-containing alpha tocopherol acetate failed to enhance in vivo killing of B. abortus.

Abstract: In the sero-epidemiological study in Union Territory of Delhi. sera from 148 veterinary workers and fifty healthy individuals were examined. Of the serum samples collected from veterinary workers 41 (27.7%) showed a titre of 80.I.U./ml for brucella agglutinins. Among positive individuals 9 (22%) exhibited clinical features. The highest sero-positivity was recorded in veterinary compounders (51.4%) followed by veterinary assistant surgeons (40%). Agewise positivity was found to be maximum (37.3%) in the age group 31 to 40 years and minimum (11.6%) in the age group 21 to 30 years. The incidence was found to be higher in rural than urban districts. In healthy control group none was found to be positive.

Abstract: The incorporation of B. abortus protective antigen into liposomes and its localization in liposomes have been found to depend on the lipid composition of liposomes. After the injection of the protective antigen conjugated with negatively charged liposomes humoral response is more pronounced than after the injection of the protective antigen incorporated into neutral liposomes. The immunization of guinea pigs with antigen-containing liposomes ensures the production of "incomplete antibodies" in the animals in high titers.

Flores-Castro R, De Cordoba LF, Trejo-Salomon J, Del Rio-Vargas J. Adult cattle vaccination and revaccination with strain 19 reduced doses for the control of brucellosis: a field experience in Mexico. International journal of zoonoses. 1985 Dec; 12(4); 299-303. [PubMed: 3939134].

Abstract: The scientific and organizational development of an effective prophylaxis against infections in animal husbandry results from the fact that many zoonoses, like salmonellosis, campylobacteriosis, toxoplasmosis, leptospirosis, listeriosis, rickettsiosis (Q-Fever) and cysticercosis as well as certain important virus infections with regard to meat hygiene cannot be detected during official ante- and postmortem inspection. The cause of these infections is clinically inapparent and leaves no pathologic-anatomical lesions. Partly responsible for these latent infections is mass production with its specific forms of husbandry, particularly in poultry and pigs. The development of these animal production methods as well as the spread of the aforementioned zoonoses in man and animal is being discussed in this paper. The information on zoonoses is based on cases reported in accordance with the Federal Communicable Diseases Act and/or the regulations on notifiable animal diseases. The potential harmfulness to the consumer's health, especially in view of his food habits, is discussed in the light of the increase of foodborne infections and intoxications caused by Salmonella. Up until now, several regulations exist to keep causative agents of zoonoses away from animal farms. In view of the successful eradication of tuberculosis in cattle and brucellosis, it is recommended on a longterm basis, to eliminate those zoonoses from animal farms, which are of special importance from the meat-hygienic point of view. On a medium-term basis, examination of farm animals should be introduced voluntarily prior to the official ante- and postmortem inspection. It is of vital importance to establish the necessary diagnostic and practical conditions for the herd-tests. A recommendation worked out by the European Community for the examination of broiler-farms is welcomed as an example of prophylactic measures suitable for the improvement of consumer protection.

Abstract: An epidemiological survey was conducted after the observation of 4 cases of acute brucellosis in an horticultural school. Of a total number of 215 attendants, eventually exposed to a single contamination, 65 cases of brucellosis were thus identified. The source of contamination was probably a practical lesson where the pupils studied a bovine gravid uterus. More stringent regulations are needed in order to avoid further similar epidemics.

Abstract: Human brucellosis in Florida is documented for the 47-year period 1928-1975. Of the 936 cases reported in 1930-1975, more than half (505) occurred in the decade 1940-1949. The incidence declined rapidly to an average rate of five cases per year, although there was an increase in 1974 and 1975. The analysis is mainly concerned with the years 1961-1975, since more complete epidemiologic data are available for this period. Cases occurred throughout the year, with the highest incidence being between May and August. White males in the 25-35 and 35-44 deciles were most affected. Of occupations identified between 1963-1975, 44% were related to the livestock-producing industry, 16% to the meat processing industry and 28% to hunters, housewives, students and children. Twelve per cent of the cases did not fall in any of these categories. Cattle were the most probable source of human infection in the livestock industry, and swine were associated with most non-industry cases. In 1963-1975, B. suis was isolated from 10 cases and B. canis from one case; one other isolate was not identified beyond genus. Of the 27 human cases recorded during 1974-1975, 15 (56%) involved swine contact (five livestock industry, four consumers and six hunters), 11 (41%) involved cattle contact (all from the livestock and meat packing industries), and one involved contact (with both cattle and swine (livestock industry). Human cases contracted from swine were scattered statewide, whereas cases associated with cattle only occurred in counties that had )( high densities of cattle and 2) modified-certified ratings in the Brucellosis Eradication Program.

Abstract: Variations in Brucella milk ring test reactions have been attributed to size and disparity in size of fat globules, fat content of milk, inhibitory factors, and cream-rising capacity. Physical properties of milk fat globules were studied in individual milk samples that had diverse sensitivities for detection of Brucella agglutinins. Size or disparity of globule size could not be correlated with the milk ring test sensitivity. Inhibitory and enhancing creaming factors could be transferred from the cream to the skim milk and reacted with other cream to show changes in sensitivity. The proportion of clustered milk fat globules was related directly to the quantity of agglutinins detected by the test. Sensitivity of the test was attributed to a fat globule agglutinin that caused clustering and to an inhibitory factor.

Abstract: A study was undertaken to compare several enrichment and direct isolation media for their suitability to detect and enumerate five strains of Campylobacter jejuni in refrigerated (5 degrees C) chicken meat. The influence of CO2 on survival at 5 degrees C was also investigated. Selective enrichment media evaluated included Preston broth (PB), selective semisolid brucella medium (SSBM), Campylobacter enrichment broth (CEB), VTP brucella-FBP broth (VTP), Rosef and Kapperud Campylobacter enrichment broth (RKCEB), and Doyle and Roman enrichment broth (DREB). Direct isolation agars included Campy brucella agar (CBAP), blood-free Campylobacter medium (BFCM) and modified Butzler agar (MBA). Comminuted chicken meat was inoculated with C. jejuni, sealed under atmospheric gas or CO2, and stored at 5 degrees C for up to 21 days. Viable population was determined by the most-probable-number technique (PB, SSBM, CEB, VTP, and RKCEB, followed by plating on CBAP, BFCM, and MBA), enrichment on DREB, followed by plating on CBAP, BFCM, and MBA, and direct isolation on CBAP, BFCM, and MBA. Without exception, direct plating of samples was superior to the most-probable-number technique for enumerating C. jejuni; MBA was inferior to CBAP and BFCM, and DREB performed at least as well as other enrichment media evaluated. Carbon dioxide afforded protection against death of three of the five strains of C. jejuni tested.

Abstract: The effect of oral administration of brewer's yeast on resistance to infectious diseases was studied in laboratory animals. It was shown that there was a significantly increased enhancement of resistance to seasonal respiratory and enteric infections in rhesus monkeys. Similarly, enhanced resistance to experimental chronic infections was observed in mice after yeast administration. A 2-week lag occurred between the initiation of yeast treatment and the expression of enhanced resistance. Study of the mechanism of the yeast-induced enhancement of resistance to infection leads to the conclusion that it is based on in vivo stimulation of phagocytosis, as measured by the "phagocytic index." No effect of brewer's yeast on circulating antibody levels was detected.

Abstract: With an L1210 tumor vaccine model, three biological and two chemical agents were tested for their ability to act as adjuvants. Adjuvant was administered with irradiated L1210 cells to immunize mice against this poorly immunogenic tumor. Two chemicals, pyran copolymer and glucan, and one biological, Brucella abortus strain 456 ether extract, were shown to be strong stimulators of antitumor immunity. Vaccination with irradiated tumor cells or adjuvant alone did not produce host resistance. Optimal immunity to challenge was produced by concomitant administration of either pyran copolymer, glucan, or B. abortus strain 456 ether extract with L1210 vaccine. Antitumor immunity was maximally expressed when vaccine and adjuvant were administered i.p. Evidence for systemic immunity was demonstrated when challenge was at a distal s.c. site. Mice immune to challenge were found to be refractory to a later rechallenge.

Abstract: The Milk Ring Test (MRT) can be used to detect Brucella antibodies in tank milk if the following points are considered: (1) Before testing, the milk must be stored at least 72 hours at 6 degrees C. (2) To obtain a reliable reaction one must use 8 ml milk and 0.8 ml antigen, instead of 1 ml milk and 0,05 ml antigen. (3) The 4 vol.% antigen appears to be more sensitive than the 5 vol.% antigen. (4) A standard negative milk sample should be included in a batch to be examined. It can help to determine doubtful results. (5) For further determination of a doubtful result obtained from the 1 ml milk reaction, 3 ml milk and 0.05 ml antigen can be used.

Garcia MM. Evaluation of various media and the use of soil infusion in the production of Brucella abortus antigen. Canadian journal of comparative medicine. Revue canadienne de medecine comparee. 1973 Jan; 37(1); 33-42. [PubMed: 4265551].

Abstract: Brucella abortus (ADRI 413) was grown on various synthetic, commercial, laboratory-formulated and soil infusion-supplemented agar media. Of 22 media tested, three soil infusion-supplemented media and a serum dextrose agar yielded more brucella antigen than the meat infusion agar, the standard reference medium. Results from agglutination and stability tests indicated that antigens produced from most of the laboratory-formulated media and the soil infusion-supplemented agars compared favourably with that derived from the meat infusion agar. The presence of meat infusion or commercial beef extract appeared to be a basic requirement for producing satisfactory and stable antigen. The growth enhancing effect of certain soil infusions seemed to be associated with the soil organic matter fraction. A significant correlation (p>.05) was obtained between the yield of brucella antigen and the glutamic acid content of the soil infusions. With proper screening, media containing, among other components, beef extract and soil infusions may be of value particularly in areas where the limited supply of fresh meat would make meat infusion agar uneconomical in the production of B. abortus (ADRI 413) antigen.

Vargues R. The use of the autoanalyzer for the automatic titration of antigenic preparations by means of complement-fixation. Annals of the New York Academy of Sciences. 1965 Nov 9; 130(2); 819-26. [PubMed: 4286047].

. The hygiene and marketing of fresh cream as assessed by the methylene blue test. A report by a Working Party to the Director of the Public Health Laboratory Service. The Journal of hygiene. 1971 Jun; 69(2); 155-68. [PubMed: 4326247].

Abstract: For the first time a survey of the prevalence of bovine brucellosis in three regions of the Democratic Republic of Somalia has been carried out. Blood sera were tested by the SAT and CFT and milk samples by the MRT. An average prevalence rate of 9.5 per cent (SAT) and 12.0 per cent (MRT) was found. There was considerable differences between regions and the reasons for this are discussed.

Farrell ID, Robertson L. A comparison of various selective media, including a new selective medium for the isolation of brucellae from milk. The Journal of applied bacteriology. 1972 Dec; 35(4); 625-30. [PubMed: 4631240].

Abstract: Chickens irradiated and bursectomized in the newly hatched period consistently develop agammaglobulinemia and form no circulating antibodies; if the birds are treated immediately after operations by intra-abdominal injection of unirradiated autologous bursa cells, immunoglobulin production, lymphoid germinal centers, and plasma cells are restored; however, the birds fail to produce antibody to specific antigenic challenge.

Gerzymisch J, Hock A, Strey J. [Studies of the different effect of the quantity and quality of dietary proteins as well as of amino acid supplements on the growth and antibody formation against Brucella in weaned rats. 2]. Archiv fur experimentelle Veterinarmedizin. 1968; 22(1); 13-23. [PubMed: 4971488].

Abstract: A case of brucellosis due to Brucella abortus, biotype 5, occurred in a bulk milk-tanker driver who collected milk from refrigerated tanks on 23 farms.The organism was isolated from blood cultures by using a serum dextrose broth containing antibiotics. Serological investigations indicate that both abortus and melitensis antigen suspensions should be used in the investigation of cases of suspected brucellosis.As Brucella abortus is an intracellular parasite heavy, prolonged, antibiotic therapy is necessary in the treatment of the disease. The danger of inadequate treatment of the acute disease is that it may become chronic, and response to antibiotic therapy in chronic brucellosis is not good.

Evans FW. Progress in eradication of bovine tuberculosis and brucellosis in New South Wales and the efficacy of a trace back system. Australian veterinary journal. 1972 Apr; 48(4); 156-61. [PubMed: 5082481].

Abstract: NA

Ebadi A. Comparison of various serological tests on the milk of sheep in relation to the isolation of Brucella melitensis. The British veterinary journal. 1971 Mar; 127(5); 105-12. [PubMed: 5102236].

Abstract: NA

Nandgoankar D, Narayana Rao PL. A survey of the incidence of bovine brucellosis in the integrated milk project area and in some of the dairy farms of Andhra Pradesh. The Indian veterinary journal. 1971 Jan; 48(1); 12-8. [PubMed: 5103570].

Leech FB. A critique of the methods and results of the British national surveys of disease in farm animals. I. Discussion of the surveys. The British veterinary journal. 1971 Nov; 127(11); 511-22. [PubMed: 5167824].

McDevitt DG. The relevance of the anti-human globulin (Coombs) test and the complement-fixation test in the diagnosis of brucellosis. The Journal of hygiene. 1970 Jun; 68(2); 173-87. [PubMed: 5270201].

Brochart M, Fayet JC, Barnouin J. [The control of animal diseases in france during the last fifteen years, as it appears throught the eradication of bovine tuberculosis. proposals for the future (author's transl)]. Annales de recherches veterinaires. Annals of veterinary research. 1979; 10(4); 575-92. [PubMed: 547830].

Abstract: In the general outline of the success, on a national scale, of the eradication of bovine tuberculosis in France up to 1974, we have tried to find out the factors which could explain the differences, sometimes important, between French departments in the efficiency of this eradication. We have selected twenty seven parameters distinctive of each of the 74 departments having more than 50 000 bovine under the control of State Veterinary service. These parameters describe 1) tuberculosis infection rates of animals and herds, at three periods; 2) some structural characteristics of bovine production; 3) sanitary environment (veterinary practicioners, State Veterinary service, breeders association for sanitary protection). Statistical analysis by principal components show that department infection rate decreases as the first three factors evidenced by this analysis vary positively; they are: 1) "Sanitary Consciousness", which is defined both by the important of bovine production and by precociousness in developing prophylactic measures, these two factors being independent. 2) Specialization in bovine production, characterized by the percentage of young animals reared in the herd and by the proportion of grass produced by ley. 3) Intensity of sanitary environment. These three factors, which are interlinked to various degrees, control respectively 25%, 13% and 13% of the total variance. A complementary analysis of the data suggest that the efficiency of eradication could be improved by a better knowledge of the movement of breeding animals between the departments, and by a more careful recording, in some instances, of infected animals in slaughter hourses. When the importance of a herd increase sanitation of tuberculosis in infected herds is less readily achieved and brucellosis abortion rate increases. At the departement level, efficiency of eradication is not correlated to the expenses of prophylaxy. For the development of an integrated sanitary policy, taking into account such non specific factors as those exemplified in the study could help to a better control of animal diseases, in parallel to the desirable improvement of the specific means of combatting each disease.

Thrower WR. Agriculture and the public health. The Milroy lectures, 1970. Journal of the Royal College of Physicians of London. 1970 Jul; 4(4); 277-304. [PubMed: 5535790].

Abstract: NA

Pospisil M. Use of Carbowax as a high molecular weight nonprotein substance for cultivation of lymphoid cells and study of their morphology. Folia microbiologica. 1967; 12(4); 367-71. [PubMed: 5593334].

Abstract: Principles of epidemiological and epizootological investigations in a brucellosis focus are presented in this work. A founded determination of a focus and methodological approaches to its study and systematization of the data concerning all the foci of a definite administrative territory is given. Also approaches to the realization of out-patient observation over the population in the brucellosis foci are characterized. The author believes that only such scientifically founded and all-round approach to the study of epizootological and epidemiological regularities of the brucellosis foci existence would aid successful solution of the most important problems pertinent to reduction of brucellosis incidence among humans.

. Brucellosis and "untreated" milk. A suggested scheme for the surveillance of producer retailer herds and the placing and removal of heat treatment notices. Monthly bulletin of the Ministry of Health and the Public Health Laboratory Service. 1967 Dec; 26; 228-30. [PubMed: 5630776].

Mathur TN. A case of brucellosis due to consumption of improperly pasteurised milk. The danger of infection with Brucella melitensis. Indian journal of medical sciences. 1968 Feb; 22(2); 106-7. [PubMed: 5694652].

Abstract: During treatment of brucellosis with sulfisoxazole, tetracycline, and streptomycin sulfate, severe thrombocytopenic purpura developed in a young farmer. Verification for an immune mechanism was provided by clinical challenge with a small dose of sulfisoxazole that caused recurrence of thrombocytopenia and by serologic laboratory test results that detected a serum factor causing platelet agglutination requiring the presence of sulfisoxazole. The original antigenic stimulation was considered to come from drinking cows' milk contaminated with sulfonamide drugs. Cross-reactivity with some other sulfonamide drugs was demonstrated.

Abstract: A three-stage sampling survey among farmers and their families living on farms in the department of Ille-et-Vilaine gave the following results: Among 490 persons examined 313 (64 per cent) showed a positive skin test and 105 (21 per cent) a positive serological reaction; 88 of the 105 patients ignored their health status, although 45 of these presented clinical symptoms (nine per cent). This survey is continuing in order to study the non-respondents and to better analyse the epidemiological situation at a farm level.

Abstract: On partition in aqueous polymer two-phase systems containing dextran, poly(ethyleneglycol) (PEG), and PEG substituted with charged or hydrophobic groups "inagglutinable" ox erythrocytes showed more negative surface charge and less hydrophobicity than "agglutinable" ox erythrocytes. Tryspin treatment of the erythrocytes increased the agglutinability, reduced the negative charge, and seemed to increase the liability to hydrophobic interaction. Haemagglutination of highly negatively charged ox erythrocytes is almost impossible to accomplish by antibody against the ox erythrocytes alone or against IgG antigen linked to the erythrocytes in passive haemagglutination. Likewise, reverse passive (antiglobulin) haemagglutination cannot be accomplished with these cells when immunoglobulin antigens of moderate molecular size (less than or equal to 900,000) are used. However, these cells may be agglutinated when a non-charged carrier such as a bacterium coated with sensitizing antibody (immunoglobulin) is used to bridge the antiglobulin-coupled erythrocytes in a mixed latticer aggllutinate. Such a bridging can also be accomplished by heat aggregation of the immunoglobulin antigen.

Abstract: Spleen cells from adult CBA/H or CBA/N mice, or from neonatal CBA/H mice, were fractionated on thin layers of fluorescein (FLU)-gelatin to yield FLU-specific B lymphocytes. A single cell, or small numbers ranging from 1 to 10, were cultured in 10-microliter microcultures together with various antigens and mitogens. The results were compared with those of bulk culture or limiting dilution cultures supported by thymus filler cells. B cell growth and differentiation-promoting conditioned media (BGDA) were added to some cultures. The CBA/N results gave no support to the commonly used classification of T cell-independent (TI) antigens into TI-1 and TI-2 categories. A typical supposed TI-1 antigen, FLU-LPS, strongly stimulated normal adult single FLU-specific B cells to proliferate and form antibody, but virtually failed to trigger CBA/N B cells of comparable antigen-binding avidity. The same was true of LPS or LPS plus dextran sulfate acting as mitogens. The allegedly TI-2 antigen FLU-Ficoll, although still triggering comparatively poor responses, was actually marginally more active than FLU-LPS. FLU-Brucella abortus (FLU-BA) + BGDA gave the best results with single CBA/N B cells, but still induced only 1.27% of cells to develop into antibody-forming clones vs 12.2% with CBA/H cells. The results obtained with single neonatal B cells also lent no support to the distinction between TI-1 and TI-2. Both "TI-1" and "TI-2" stimuli caused adequate proliferation, one "TI-2" antigen stimulating 23.2% of the cells. None of the antigens caused good antibody formation, however, probably because multivalent antigens can deliver signals impeding the differentiation of immature B cells. It is therefore suggested that the classification of TI-1 antigens into two subcategories be abandoned, at least for the time being.

Abstract: Incorporation of lipid A influences the immunogenic properties of liposomal model membranes that carry N-(hapten)-substituted derivatives of phosphatidylethanolamine as antigenic determinants. These effects of lipid A include a marked enhancement of the hapten-specific response; abolition of the requirement of critical threshold epitope density for liposomal immunogenicity; and conversion of liposomes from a thymus-independent type 2 to a thymus-independent type 1 immunogen. Recent experiments demonstrate that an optimal in vitro response to liposomes--although originally classified as thymus-independent on the basis of conventional criteria--requires two T cell factors, interleukin-2 and interleukin-X, which are also involved in the response to a classic thymus-dependent immunogen. Incorporation of lipid A has no qualitative effect on the dependence of liposomal immunogenicity on these lymphokines.

Abstract: Various chemotherapeutic regimens were evaluated in 48 culture-positive dairy cows. Cessation of shedding of Brucella abortus from udder secretions and absence in selected tissues at necropsy were criteria of success. A combination of a long-acting oxytetracycline and streptomycin eliminated Brucella in 10 of 14 (71.4%) cows. Two cows that were retreated with the same regimen also became culture-negative. Other treatment regimens, including the use of liposome-encapsulated antibiotics, were less successful. Serotests were a poor criterion of effectiveness.

Abstract: This report describes the sensitisation of latex particles and the use of this reagent on the Autotape system to select blood donations containing greater than or equal to 4 IU/ml of tetanus toxoid antibody. Plasma containing tetanus toxoid antibodies is processed to provide immunoglobulin for passive protection against tetanus. The techniques used for screening include immunoelectrophoresis [3], tetanus toxoid sensitised latex particles [5], haemagglutination [1] and automated haemagglutination [6]. The technique used by the Sheffield Regional Transfusion Centre until recently, was immunoelectrophoresis, selecting donors with 4 or more IU/ml. The Autotape system, previously described for brucellosis [4] and syphilis testing [2], provides a system for automated slide testing and a 5-min tetanus toxoid coated latex test has been developed to select suitable donations for the production of specific immunoglobulin.

Abstract: The aim of the study was to elucidate the possibility of using bacteriocinogenicity of Brucella as taxonomic feature, to determine their phylogenetic relation to other microorganisms by their bacteriocinogenic properties and to investigate the physicochemical properties of brucellacin and conditions for its stable detection. The Brucella cultures were isolated in the Caucasus. Investigation of their capacity for production of bacteriocin according to the procedure described by M.A. Konstantinova and A.D. Garmazova (1979) revealed that 62.1 per cent of the 216 cultures tested produced brucellacin. Isolation of bacteriocin with the methods developed was shown possible in all of the tested strains of B. melitensis, B. abortus, B. suis and in most of the strains of B. ovis. The methods also provided an increase in the synthesis and activity of brucellacin. The analysis of the characteristic features of bacteriocinogenicity and the properties of bacteriocin allowed recommending the use of additional taxonomic features for identification and differentiation of Brucella. Sensitivity of the indicator strains of Brucella to bacteriocins of other species (F. tularensis, Campylobacter fetus intestinalis B-8833, Y. enterocolitica, Vibrio cholerae and E. coli Fredericq) was noted which was additional evidence of the phylogenic relation between the above organisms. Investigation of the physicochemical properties of brucellacin confirmed the suggestion of the protein nature of the active principle of brucellacin and its similarity in different Brucella species.

Abstract: The outer membrane of Brucella species contains two major proteins, denoted as group 2 and group 3 proteins (Verstreate et al., Infect. Immun. 35:979-989, 1982). We reconstituted proteoliposomes from the purified proteins and egg phosphatidylcholine and showed that group 2 proteins, but not a group 3 protein, had the porin activity. The influx rates of sugars of various sizes into the proteoliposomes indicated that the porin channels had apparent diameters in a range comparable to that of Escherichia coli OmpF porin and that the channels were predominantly open. Among different Brucella species, there were small but detectable differences in the channel diameter, and it was possible to explain the differential sensitivity of several Brucella species to diagnostic dyes on the basis of these observed differences.

Abstract: Despite advances in the control of the disease in animals, brucellosis is a zoonosis with important economic impact in the United States. It is estimated that cases of brucellosis in humans are underdiagnosed and underreported and that the disease continues to be a major human health hazard. Nine active cases of brucellosis in humans were documented in Houston, Texas between June 1981 and June 1982. These cases, plus an additional case of chronic brucellosis, served as the impetus for this review of the protean manifestations of the disease in humans.

Abstract: Enzyme-linked immunosorbent assays were conducted on milk of cows from which Brucella abortus was isolated and that of noninfected controls. Horseradish peroxidase-labeled rabbit antibovine immunoglobulins IgG, IgG1, and IgA were used as conjugates. A heat-killed whole-cell suspension of B abortus strain 19 was used as the antigen. Differences in antibody profiles were observed in milk of cows from which B abortus was isolated and in milk of noninfected cows. Antibody profiles were similar in milk of cows infected with B abortus and that of cows from which B abortus strain 19 was isolated.

Abstract: Various buffers (0.2 M acetate, pH 4.2; 0.01 M phosphate with 0.15 M sodium chloride, pH 7.2; 0.15 M borate, pH 8.2 and 0.025 M carbonate, pH 9.6) were used as coating buffers with Brucella abortus alkali treated lipopolysaccharide on two types of plastic matrices. Maximum binding occurred using the phosphate buffer. However, as was the case with the other buffers 50 per cent or more of the antigen was removed by five washing procedures. B abortus S19 or S2308, suspended in ammonium acetate-carbonate buffer, pH 8.2, was shown to bind maximally when 10(9) cells were dried onto the plastics. Less than 20 per cent of the cells were removed by five washings.

Abstract: In the past zoonoses that caused serious human illness also caused serious loss of animal production, but there is growing awareness of the public health problems arising from infections that cause little or no such loss. Much can be learnt from the history of the control of bovine tuberculosis and brucellosis. In both cases there was reluctance to accept that animals were the principal cause of infection, and the earliest attempts at control failed because measures were taken only against clinical cases of the disease. The essential features in control of both infections were: official recognition of a problem, willingness of governments to allocate resources, and cooperation between the medical and veterinary professions. Salmonellosis is the most important zoonotic infection in Britain today, though several Orders have reduced the reservoir of infection in food animals. It is suggested that a national team of doctors should be set up to investigate and control zoonoses, that this team should be answerable to a central agency, and that it should build up close working relationships with the nominated officers of the veterinary profession.

Abstract: An indirect sandwich enzyme-linked immunosorbent assay, using antibody covalently coupled to nylon beads, has been adapted for the detection of Brucella antigens. Optimum conditions were achieved by incubation of 1 ml of reaction mixture with a single bead, and by minimizing nonspecific interactions through the use of beads coated with purified bovine antibodies, preabsorption of third layer rabbit antibodies with normal bovine serum, and treatment of beads with normal goat serum before addition of the goat anti-rabbit enzyme conjugate. Beta-galactosidase was selected for use with clinical samples primarily because of low levels of endogenous enzyme in bovine leukocytes. Use of a fluorogenic substrate enhanced sensitivity 20-fold. Under these conditions, 100 fg of solubilized crude lipopolysaccharide or 8 to 10 Brucella cells was detectable in a fixed volume of 1 ml. A system was also devised for concentrating antigen which permitted ready detection of 2 pg of lipopolysaccharide in a volume of 50 ml (40 fg/ml). Attempts to detect lipopolysaccharide in the presence of concentrated serum or plasma were unsuccessful, but 10 brucellae added to a suspension of leukocytes from 100 ml of normal bovine blood were easily measured.

Abstract: Colostrum or early breast milk or both from each of 16 healthy women contained agglutinating antibodies for all normal streptococcal inhabitants of the human oral cavity (S. mutans, S. sanguis, S. mitis, and S. salivarius), including those which colonize the neonatal oral cavity in significant numbers. Agglutination correlated with the amount of immunoglobulin A (IgA) binding to bacterial surfaces as measured by mixed reverse passive antiglobulin hemagglutination. Surprisingly, colostral IgA agglutinated our control organism, Brucella abortus. Low levels of colostral or milk IgM and IgG antibodies also reacted with all of the test bacteria. Absorption studies with an enzyme-linked immunosorbent assay showed that a proportion of antibodies in colostrum and early milk is specific for each of the different oral streptococci. Fractionation on Sepharose 4B indicated that 11S secretory IgA is the predominant form of colostral and milk antibody for all of the test bacteria, including B. abortus. No evidence was found that reactions other than antigen-antibody reactions resulted in binding of colostral immunoglobulins by any of the test bacteria.

Abstract: A total of 1246 animals (778 sheep and 468 goats) were examined for detection of brucellosis using serum tube agglutination test (SAT), complement fixation test (CFT) as well as Abortus Bang Ring test (ABR-test) have been conducted on both dairy ewes and goats. Eighteen ovine sera played positive serum agglutination reactions (2.31%). Out of the total 468 goat's serum samples examined, 22 (4.70%) gave a positive titers and 2 (0.43%) gave doubtful reactions. The reactors of sheep to the CFT was 2.44% and the incidence was lower in rams (1.63%) than among ewes (2.97%). Seven out of the 339 ewe's milk samples were positive to ABR-test (2.06%), 7 were doubtful (2.06%). Out of 354 goat's milk samples examined, 14 were positive to ABR-test (3.96%). CFT was considered to be a reliable procedure for detecting brucella infected animals particularly in male more than the SAT. Among dairy ewes, both tests were identical in their results.

Abstract: Data for path analysis on the epidemiology of bovine brucellosis were gathered from 184 premises in Mexicali Valley, Mexico, that had dairy cattle, swine, and/or goats. Each farmer manager or owner was interviewed, using a pretested set of questions, and the livestock on the premises were serotested for Brucella antibodies. The focus in gathering these data was to identify the characteristics of premises with seropositive cattle in comparison with characteristics of premises with no seropositive cattle.

Abstract: A total of 5059 sera were collected from adult female beef cattle at Cato Ridge Abattoir and tested for Brucella abortus antibody levels over the period November, 1981 to August, 1982. The sera were screened using the Rose Bengal Plate Test, and the Complement Fixation Test was used as the definitive test. Seventy-seven sera (1,5%) had titres of 30 IU complement fixing B. abortus antibody per ml serum or greater.

Abstract: The world literature was searched to document the variables known to affect the initiation, spread, maintenance, and/or control of bovine brucellosis. Each variable was classified into 1 of 3 categories, depending upon whether it was related to the animal population, to management, or to the biology of the disease. These variables were documented and categorized as the initial step for a quantitative path analysis on the epidemiology of bovine brucellosis in a defined geographic area in northern Mexico.

Abstract: Twenty-eight known or hypothesized variables in the epidemiology of bovine brucellosis were examined by path analysis to determine the contribution of each variable to seropositivity of cattle in Mexicali, Mexico. The variables that contributed significantly to bovine seropositivity were area size of farm premises, percentage of animals on a premises that were inseminated artificially, local marketing of the products from a premises, size of investment in the livestock, whether or not dairying was the major agricultural activity of the premises, and the policy of the owner or manager with regard to disposal of reactor animals. The contributions of all other variables were insignificant.

Abstract: Brucellosis constitutes a true public health problem in Upper-Corsica. The purpose of this epidemiologic study carried out in the department was to better determine the parameters of this endemic disease in order, first, to plan--and, later, to evaluate--the prophylactic measures adopted by the authorities concerned. The methodology was based upon the number of cases reported by general practitioners and by directors of pathology laboratories. The major result was to establish the fact that the disease very widely affects the general population. The majority of cases recorded are among urban dwellers, indirectly contaminated through the consumption of local, unsterilized, dairy products. Thus, preventive measures should concentrate principally on promoting improved inspection of the products concerned and on public health education.

Abstract: Lots of five weeks old female CD-1 mice were vaccinated at several doses with living (B. abortus 19), killed whole cells or purified fractions vaccines. The doses were the number of bacterial cells injected per mouse, either viable (living vaccines), total (killed) or number calculated from the yield of extraction. The mice were intraperitoneally injected with a lyophilized standard challenge, 2 X 10(5) Brucella abortus 544, thirty days after vaccination. In two assays, B. melitensis H.38 was used as challenge at several doses. Responses to a good live vaccine were dose independent. Responses to killed whole cell and purified fractions were usually dose dependent. A reference lyophilized vaccine previously prepared and titrated would give the reference response to one unit. These data can be used to devise a general control method of activity for anti-Brucella vaccines. This was proposed and discussed in the preceding paper.

de Reviers M, Fensterbank R. [Biological control of the brucellin INRA in sensitized guinea pigs: a guide for the statistical analysis of results]. Annales de recherches veterinaires. Annals of veterinary research. 1984; 15(3); 395-403. [PubMed: 6517480].

Abstract: Brucellin INRA is a new allergen used for the screening of brucellosis. It shows, by a delayed-type skin hypersensitivity reaction, the sensitization induced in animals by a brucella infection. The biological potency of each batch must be compared to that of a standard allergen. The use of guinea-pigs necessary for this quality control and the statistical analysis of the results are described and discussed. The aim of this paper is to provide the end-user with a simple and accurate method, and to alert him to the difficulties and limits of such a test.

Abstract: Of 4,144 serum samples collected from cows on 113 farms from eight areas of Colombia 3.3% had positive and 8.8% inconclusive titres to Brucella abortus, 21.7, 6.3, 1.6, 0.6 and 0.7% of cows had positive titres to Leptospira serovars hardjo, pomona, canicola, icterohaemorrhagiae and grippotyphosa respectively. Questionnaires completed on 110 farms revealed that 6, 2.5 and 4.6% of cows had had metritis, aborted or retained their placentas respectively in the previous 12 months. Trichomonas foetus and Campylobacter fetus were isolated from 13.7% and 15% of the bulls sampled on 103 farms. Six and two bulls had inconclusive and positive titres to Brucella abortus. Eight and 23 bulls had positive titres to pomona and hardjo. The results were discussed and remedies for control suggested.

Abstract: IgA, IgM and IgG concentrations and their bacterial antibodies to E. coli, group B streptococci and Brucella abortus were measured in human breast milk collected from the 1st to 10th day post-partum from mothers delivered of 'preterm' infants (Premature Breast Milk or PBM) and from mothers delivered of term infants (Term Breast Milk or TBM). Reverse passive haemagglutination tests (RPH), rocket immuno-electrophoresis and mixed reverse passive antiglobulin haemagglutination tests (MRPAH) were employed. PBM at 2-5 days post-partum (though not beyond this period) contained higher IgA levels than did TBM, and this difference persisted even when total IgA was expressed as a proportion of total milk protein: in contrast the IgM and IgG contents of PBM and TBM were the same at both these postnatal ages. The titre of IgA antibody to E. coli, which was absorbable only by the corresponding bacteria, showed no significant difference between PBM and TBM, whereas the titres of IgA reacting with Br. abortus and, to a lesser extent group B streptococci, were higher in PBM than those in TBM. However the IgA which reacted with Br. abortus and group B streptococci was not specific to those organisms but was absorbed by all three bacteria studied. It is speculated that the high IgA content of early preterm milk and perhaps the presence of especially high titres of what appears to be a non-specific or cross-reacting bacterial IgA in such milk, may be immunologically advantageous to low birthweight infants fed on their own mother's milk.

Abstract: Estrual cows are more likely to be recognized if penned with other open cows. Cycling cows fresh less than 100 days, without signs of heat or a follicle but with good uterine tone and signs of ovarian activity, are closely observed for heat 15-20 days after examination. Those fresh at least 150 days are given prostaglandin. Lugol's iodine solution is infused into the uterus of cows without signs of ovarian activity but with normal uterine tone. Cows with retained fetal membranes are given an intrauterine infusion of oxytetracycline with the membranes being removed only if it will not damage the uterus. Oxytetracycline or chlorhexidine solution is infused into the uterus of cows with a purulent vulvar discharge 2 or more weeks postpartum. Prostaglandins may benefit cows not showing signs of heat by 21 days postpartum. Procaine penicillin G solution is infused if the uterus is abnormal at 60 days postpartum. Heifers are vaccinated against brucellosis, blackleg, leptospirosis, IBR-PI3 and BVD at 4-11 months of age, with an optional Vibrio vaccination if a bull is used in the herd. Cows are given IBR-PI3, BVD and Leptospira vaccines a few days post-partum, with a Leptospira booster at pregnancy examination. Cows with Streptococcus mastitis in late lactation may best be treated by drying off. Control of coliform mastitis necessitates a clean barn and milking parlor environment. Staphylococcus mastitis may require segregation of affected cows and installation of a low-line milker to prevent spread.

Abstract: The present three-year longitudinal study has clearly demonstrated the advantages of modern and scientific livestock management over the traditional and extensive methods of animal husbandry in the prevention and control of bovine and human Brucella abortus infection. The occupational hazard, in relation to brucellosis, to which livestock farmers are exposed in Oyo State is closely related to the level of infection in the animal population, the level of environmental contamination and hygiene on the farm. This study has also revealed that many farmers could acquire subclinical brucellosis within two years of constant occupational exposure to source of infection. There was no seasonal variation found in the prevalence of brucella infection among the human and cattle population studied at the I.A.R.T. farm in Ibadan: climatic variation is therefore considered to play no important role in the prevalence of brucellosis in Southern Nigeria.

Abstract: During the period 1977-1978, a serological and epidemiological investigation was conducted by the Victorian Departments of Health and Agriculture to determine the risk to abattoir workers of exposure to Brucella abortus from infected cattle. Nearly 1000 specimens of serum from workers at three abattoirs were tested for brucella antibodies at the Microbiological Diagnostic Unit. Serum agglutination and antihuman globulin tests were performed by a microtitre technique. Approximately 25% of these specimens gave positive results, with a wide variation in frequency in different abattoirs. Data relating to personal, employment and medical history were obtained by means of questionnaires. An attempt was made to identify risk factors by analysis of this information and the serological results. Within each abattoir, the presence of detectable antibodies was no more likely in people with symptoms than in asymptomatic people working in the same abattoir. However, the proportion of workers with symptoms was much higher in two abattoirs with a relatively high percentage of seropositive workers, than in a third, in which the percentage was comparatively low. There was evidence that the risk of exposure to B. abortus correlated with the percentage of cows in the total yearly "kill" for each abattoir.

Abstract: The possibility of brucellosis should be considered in any case of obscure pyrexia, monoarthritis, fascitis or splenomegaly, especially in persons who have been in contact with cattle, pigs or goats, or who have recently visited the Mediterranean region or East Africa. Tetracycline controls the symptoms, and the addition of streptomycin reduces the tendency to relapse.

Abstract: The prevalence and persistence of Brucella abortus strain 19 shedding from the udder were studied in adult cattle in infected dairy herds that were vaccinated by 4 methods. Specimens were cultured from 1,736 cows in 55 vaccinated herds. Of the 744 brucella isolates, 98 were strain 19 and were from cattle in 25 of the herds. Shedding was highest among conjunctival vaccinates (1.4%) and higher in those given the standard subcutaneous dose (0.83%) than in those given a reduced subcutaneous dose (0.45%). Isolations were not made from intradermally vaccinated cattle. The persistence of strain 19 shedding was related to the length of time after the initial isolation. Approximately 20% of the shedders that remained in the herds were seropositive 13-18 months later. Cattle were selected for bacteriologic studies largely on positive results of rivanol or complement-fixation tests inasmuch as the card and tube agglutination tests were unreliable. Of the 646 field strain isolates, 95.8% were biotype 1, 2.5% were biotype 2, and 1.7% were biotype 4.

Abstract: An enzyme-linked immunosorbent assay (ELISA) method was evaluated for the detection of antibodies to Brucella abortus in cows milk. Milk samples from seropositive or -negative cows were sed to determine the distribution of absorbance values to classify milk as ELISA positive or ELISA negative. Brucella abortus was isolated from milk samples from 10 (45%) of the 22 cows whose milk and serum were ELISA positive. The ELISA was evaluated and determined to be an appropriate method for detecting antibodies to B abortus in bovine milk.

Abstract: Three clinically normal cows were inoculated with viable Yersinia enterocolitica O9 by the intramammary route. This provoked an increase in the somatic cell count in milk from the inoculated quarter, but no obvious clinical response. Yersinia enterocolitica was isolated only sporadically from these cattle after inoculation. All cows showed a marked serologic response to Y enterocolitica and Brucella abortus. The anti-Brucella response provoked by inoculation of Yersinia was sufficient to render milk and serum Brucella-seropositive, as measured by the standard milk ring test, the quantitative milk ring test, and the serum agglutination test.

Abstract: A process of emulsifying and centrifuging abattoir specimens before plating out is described. Brucella abortus was isolated more successfully by this process than by conventional methods, especially in low grade infections. The 5 different media used were equally effective in our attempts at isolation, but growth did not necessarily occur on all 5 plates. In dairy cows, specimens from supra-mammary lymph nodes, udder and iliac lymph nodes accounted for a high percentage of positive isolations.

Abstract: An indirect enzyme-antibody immunosorbent assay (BrucELISA) is described for the detection of antibody to Brucella abortus in cow's milk. Three series of milk samples were obtained from an adult-vaccinated dairy herd infected with B. abortus. The BrucELISA system was used as a screening test for individual milks diluted 1:200 (BE 200 test), for undiluted bulk milks, and to determine antibody titer (BrucELISA titration assay). The BrucELISA results correlated highly with positive Brucella ring test reactions and culture positivity, eliminated false-positive Brucella ring test reactions, detected antibody in some samples which were Brucella ring test negative, and distinguished between vaccinated and infected animals. BrucELISA titration assay titers of greater than 1:800 were correlated with shedding, or were prognostic for animals which eventually became shedders. Binding of the enzyme-antibody conjugate to bovine immunoglobulin in the absence of rabbit anti-bovine immunoglobulin occurred with culture-positive or -negative milks showing titers of greater than 1:1,600 (the beta effect); the effect was also of predictive value in identifying eventual shedders. The BrucELISA system is a sensitive, specific, and inexpensive method for screening large numbers of individual or bulk milk samples for the presence of antibody to B. abortus.

Abstract: During the period October 1979 to May 1980, 22 cases of acute brucellosis occurred at a South Australian abattoir. We obtained blood samples for serological investigations and culture of Brucella abortus, and tested the sera using the standard agglutination test (SAT), dithiothreitol test (DTT), anti-human globulin test (AHG) and complement fixation test (CFT). Patients showed large variations in antibody levels in each of these tests, both on presentation and after treatment. Blood culture was successful in six of 15 patients in whom it was attempted. Using the experience gained during the outbreak, we defined a set of serological criteria for the diagnosis of acute brucellosis, particularly in those who are occupationally exposed.

Abstract: Among a number of synthetic polymers under study polyvinylpyrrolidone was found to be most active in stimulating the immunogenic action of brucellosis protective antigen. It was also found that the polymer with a molecular weight of 490,000 had a longer stimulating effect on the protective properties of brucellosis protective antigen than the polymer with a molecular weight of 40,000. The intramuscular injection of brucellosis protective antigen had no sensitizing effect on the body and produced no pronounced local and systemic reaction.

Abstract: In order to extract the polysaccharide B of brucella used in Radial Immunodiffusion test (RID) for detection of infected cattle, a quick, simple and safe method is proposed. It consists of using phenol killed cells of B. melitensis 16 M known to produce polysaccharide B. After autoclaving brucella cells resuspended in saline, the cells are separated by centrifugation and the polysaccharide B is obtained by two successive ethanol precipitations of the supernatant. Comparison of the results of RID tests carried out with the polysaccharide extract of B. melitensis 16 M (rapid method) and 231 sera of infected cattle shows the validity of this method of preparation. Differences in yield (1.5% for the second precipitate against 0.5% for the polysaccharide-reference and in optimum dose used the reaction (10 microgram against 200 microgram) show that as many as 60 times more RID tests can be made with the polysaccharide obtained by rapid method.

Abstract: In England and Wales between 1951 and 1980 233 reported outbreaks of communicable disease attributed to milk or dairy products affected nearly 10 000 people, of whom four died. Tuberculosis and brucellosis have been controlled, but milk-borne outbreaks of salmonellosis and campylobacter enteritis due to raw or defectively pasteurised milk are common and may be increasing in number. Universal heat treatment of milk is an effective preventive measure, and it is regrettable that the continued sale of untreated milk is to be permitted in England and Wales.

Abstract: Ten stabilizing media commonly used by commercial laboratories for the stabilization of Brucella abortus strain 19 for lyophilization were studied. Six media gave similar results, with viable count averages ranging from 50% to 58%. The standard stabilizer recommended by the World Health Organization (casitone 2,5%, sucrose 5% and sodium glutamate 1%) was included. The other four media gave viabilities with averages from 19,7% to 38,7%, which represented a highly significant difference. The lowest viable count was observed with the 5% polyvinylpyrrolidone stabilizer. The differences in the number of viable cells among the various processes were greater than expected.

Abstract: Cell envelopes prepared from smooth and rough strains of Brucella were characterized on the basis of lipopolysaccharide and protein content. The action of three kinds of detergents on Brucella cell envelopes and Escherichia coli control cell envelopes was examined on the basis of the proteins and lipopolysaccharides that were extracted. As compared with those of E. coli, Brucella cell envelopes were resistant to nonionic detergents. Zwittergents 312 and 316 were most effective in extracting E. coli cell envelopes, and Zwittergent 316 was most effective in extracting Brucella cell envelopes. Sarkosyl extracted proteins but extracted only trace amounts of lipopolysaccharides from cell envelopes of both bacteria. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the Sarkosyl-resistant proteins revealed a composition similar to that of the proteins exposed on the surfaces of viable cells, as determined by the lactoperoxidase-125I radioiodination method. EDTA, with either Tris-HCl or Tris-HCl-Triton X-100, did not have detectable effects on Brucella cell envelopes. Ultracentrifugation of purified lipopolysaccharides in detergents and EDTA demonstrate that, in contrast to that of E. coli, Brucella lipopolysaccharide was not stabilized by divalent cations. Sarkosyl was ineffective in dispersing lipopolysaccharides, whereas the action of Zwittergents was related to the length of their alkyl chains.

Abstract: Horses at a veterinary teaching hospital and a slaughterhouse were surveyed for antibodies to Brucella abortus, B canis and Actinobacillus equuli. Four of the 141 hospitalised horses and none of the 73 slaughtered horses had titres of 1:100 or greater to B abortus. Six horses of both populations reacted to the card test. One was culture positive. A card test using B canis antigen was positive in 38 per cent of the sera from hospitalised horses and all of the slaughtered horses. Twenty (27.4 per cent) of the latter group had high tires in a tube agglutination test. High titres could not be reduced by 2-mercaptoethanol serum treatment. The titres appeared to be associated with advanced age but not to sex. Adsorption of sera with B canis did not affect titres to A equuli but the reverse was true.

Abstract: Following concern about the recent publication indicating the possible ability of Brucella abortus to survive commercial pasteurisation of naturally contaminated milk, such milk was subjected to biological (BT), serological and bacteriological tests. The raw milk was Brucella Milk Ring Test positive and biotype I was isolated from 4/5 BT guinea pigs, the one tested being seropositive to the Rose Bengal Test, and with serum agglutination and complement fixation titres of 160 and 36 international units respectively. After batch (63 degrees C/30 min) and high temperature, short time (72 degrees C/15 sec) pasteurisation, all 10 BT guinea pigs were bacteriologically and serologically negative, indicating that officially approved methods of commercial pasteurisation rendered naturally Brucella-contaminated raw milk safe for consumption.

Abstract: A comparative study has been made of rabbit precipitating and coprecipitating anti-egg albumin antibody in rabbits repeatedly injected with soluble and particulate antigens. Four different antigens were used. a) Soluble egg albumin. b) Polymerized egg albumin. c) Egg albumin linked Brucella abortus 19 strain. d) Brucella abortus 19 strain. Rabbits were subcutaneously injected every 2 weeks during 29 weeks. Before each inoculation they were bled and the concentration of serum precipitating and coprecipitating antibodies was determined. In animals inoculated with the soluble antigen, coprecipitating antibodies were present during the whole course of the immune response and they constituted 10% of the total antibody population. In rabbits injected with particulate antigen (egg albumin linked Brucella abortus) coprecipitating antibodies were also present during the whole course of the immune response but their serum concentration was not the same in different periods of time. An increase in coprecipitating antibodies began at the 13th week reaching 50% of the total antibodies at the 28th week. Similar results had been obtained in rabbits injected with Brucella s. p. In animals inoculated with polymerized egg albumin, coprecipitating antibodies increased to high levels from the 1st to the 23rd week; during this period it constituted 50% of the total antibody population, after which, the animals became tolerogenic. On the basis of these results, the increase in coprecipitating antibodies in rabbit sera would be related to: 1) antigen physicochemical characteristics (soluble o particulate); 2) the number of epitopes in the particle, and 3) the size of the antigen particles.

Abstract: Campylobacter jejuni was isolated from 1 to 108 (0.9%) milk samples obtained from the bulk tanks of nine grade A dairy farms and from 50 of 78 (64%) cows producing grade A milk. Survival of eight Campylobacter strains in unpasteurized milk (4 degrees C) varied greatly: the most tolerant strain showed a less than 2-log10 decrease in viable cells after 14 days, and the most sensitive strain showed a greater than 6-log10 decrease after 7 days. One strain was still recoverable 21 days after the inoculation of milk. Inactivation of the different strains corresponded with an increase in milk aerobic plate count and a decrease in milk pH; however, no absolute correlation could be made between the rates of change of these parameters and the rates of campylobacter inactivation. When held at 4 degrees C, C. jejuni was most stable in brucella broth, died most rapidly in unpasteurized milk, and was inactivated at an intermediate rate in sterile milk. Our results indicate the presence and possible persistence of C. jejuni in raw grade A milk and reaffirm the need for pasteurization of milk.

Abstract: review of epidemic and endemic brucellosis at six abattoirs demonstrates a correlation between case distribution and flow of air from the kill department (stage II) to other areas within an abattoir. Air from the kill department disseminated to nearby departments led to abnormally high brucellosis attack rates for persons who worked in these areas at two abattoirs. Complete physical separation or maintaining negative air pressure in the kill department was associated with reduced risk for workers in other areas at four abattoirs. Cases in persons who had exposure to kill department air but no contact with animal tissues provide strong evidence for airborne transmission of infection. Brucellosis is also contracted through skin contact with infectious animal tissues, but this route of transmission appears less important than formerly believed.

Meyer ME, Kasten RW. Comparison of results of the automated complement fixation screening tests with conventional serologic tests in two dairy herds--one with recent and one with chronic brucellosis. Proceedings, annual meeting of the United States Animal Health Association. 1980; 84; 128-35. [PubMed: 6947284].

Abstract: We used the following criteria to determine whether liposomal model membranes are thymus-independent type 1 (TI-1) or type 2 (TI-2) immunogens: a) in vivo response of (CBA/N X BALB/c)F1 male and female mice; b) in vitro response of spleen cells from these mice; c) ontogeny of the response in cultures of (C57BL/6 X DBA/2)F1 male cells obtained from mice of different ages; d) ability of a monoclonal anti-delta antibody to block the in vitro response of C57BL/6 spleen cells. The liposomes were sensitized with N(2,4,6-trinitrophenyl-6-N-aminocaproyl)-dipalmitoylphosphatidylethanolamine (TNP-Cap-DPPE) and were prepared without and with incorporation of the B cell mitogen, lipid A. On the basis of the above criteria, liposomes prepared without lipid A can be characterized as a TI-2 immunogen and resemble TNP-Ficoll. In contrast, liposomes prepared with lipid A behave like a TI-1 immunogen such as TNP-BA (trinitrophenylated Brucella abortus). Conversion of liposomes from a TI-2 to a TI-1 immunogen by inclusion of lipid A in the bilayers was achieved under conditions of constant TNP-Cap-DPPE epitope density. Increasing the epitope density in liposomes made without lipid A does not result in transformation of immunogen type. These results, which bear on the features that may distinguish TI-1 from TI-2 immunogens, are discussed with regard to the involvement of macrophages and T cells in the response.

Abstract: The enzyme immunoassay is a highly versatile diagnostic tool whose use is rapidly spreading throughout the world. With the number of reagents, processing steps and possible protocols involved, and the growing list of devices used to automate or semiautomate the test, there is an immediate need to develop standard procedures for evaluating test performance and making diagnostic decisions. The positive and negative reference sera against which test samples are compared must be carefully selected and evaluated to insure that they are representative of field populations. Only then will it be possible to obtain uniform results within and between test facilities.

Abstract: A direct enrichment procedure was developed to selectively recover small numbers of Campylobacter jejuni, C. coli, and nalidixic acid-resistant thermophilic Campylobacter from foods. The procedure includes an enrichment medium composed of brucella broth, 7% lysed horse blood, 0.3% sodium succinate, 0.01% cysteine hydrochloride, vancomycin (15 micrograms/ml), trimethoprim (5 micrograms/ml), polymyxin B (20 IU/ml), and cycloheximide (50 micrograms/ml) that is inoculated with 10 or 25 g of food and incubated with agitation under microaerophilic conditions at 42 degrees C for 16 to 18 h. After incubation, the medium is plated directly onto Campy-BAP agar plates (M. J. Blaser et al., Ann. Intern. Med. 91:179-185, 1979), and resulting colonies that resemble Campylobacter are identified by conventional tests. The foods evaluated included raw milk, hamburger, and chicken skin which had aerobic plate counts of 10(5) to 10(9) bacteria/g. The procedure was effective in recovering as few as 0.1 cell of Campylobacter per g of food. Of the 50 isolates of Campylobacter evaluated, all were recovered from raw milk and hamburger at a level of 1 to 4 cells/g, and 41 and 40 isolaes were recovered from the hamburger and milk, respectively, at 0.1 to 0.4 cell/g. The enrichment was least effective for recovering campylobacters from chicken skin, as 7 and 26 of 50 isolates were not recovered at 1 to 4 and 0.1 to 0.4 cell/g, respectively. This new procedure is more rapid, direct, and effective than other enrichment or direct plating procedures for recovering small numbers of campylobacters from foods.

Abstract: A survey of incidences of brucellosis was carried out in 1378 lactating animals (590 buffaloes; 290 cows; 471 goats; and 108 sheep). Individual reactors to MRT were further identified with blood serum tube and quick agglutination tests.--Recorded incidences of brucellosis in the present investigation were found to be low, 1.4% in cows, 1.3% in goats, 1.2% in buffaloes and 0.9% in sheeps. The present investigation supports the wide use of MRT for screening antibodies for brucellosis in lactating animals which can be followed by SAT.--All those animals which responded to tube agglutination test were also positive for blood serum quick agglutination test.

Meyer ME. Brucellosis of dairy cattle in Libya: proposals for a control program under an intensive husbandry system. International journal of zoonoses. 1982 Dec; 9(2); 113-7. [PubMed: 7169304].

Abstract: Fusobacterium egg yolk agar is a new medium selective for Fusobacterium species and differential for Fusobacterium necrophorum. The medium is a Brucella Agar base (Difco Laboratories, Detroit, Mich.) containing vancomycin, neomycin, josamycin, and egg yolk. All species of fusobacteria grew with only minimal inhibition. The mean log difference in counts between Fusobacterium egg yolk agar and control media for 30 strains of seven species of fusobacteria was 0.1922. F. necrophorum typically showed a strong lipase reaction. Most other organisms were significantly inhibited by the medium.

Abstract: Economy of prophylactic haemoserological control of pig stocks in a district as well as of a sow breeding unit and of cattle stocks on two industrialised fattening farms has been enhanced by including blood samples from routine slaughtering. Preliminary organisational experience is reported in this paper. The samples collected samples collected from normally slaughtered selected sows may contribute to an improvement of veterinary production control.

Abstract: Twenty-two cases of pyrexial illness which occurred amongst workers in a country abattoir were investigated retrospectively for Q fever, brucellosis, and leptospirosis. In 18, the illness was shown to be Q fever. No diagnoses were established for the other four. The demonstration of circulating Q-fever-specific IgM globulin was instrumental in establishing the diagnosis in many of the cases.

Abstract: Throughout 1976--1978, 757 brucellosis cases have been found in Poland. Mostly such occupational groups have been afflicted with brucellosis as: veterinary service, cowkeepers, Bacutil workers and workers of meat plants. The greatest prevalence occurred in the following age groups: 41--50 (22.0%) and 51--60 (17.1%).

Abstract: The outbreak of human brucellosis among employees of a large South Australian abattoir described previously coincided with an increase in the number of cattle showing a positive serological reaction for Brucella abortus being slaughtered. Comparisons showed that two other abattoirs in the area were slaughtering larger numbers of such cattle, but no cases of human brucellosis were diagnosed there. This suggested an additional risk at the abattoir concerned. All infected men had been employed in a particular part of the works. There was a possibility of movement of aerosols, produced on opening the uteri of pregnant cattle, to other parts of the works, putting a larger number of workers at risk of infection. Modifications to the plant greatly reduced the spread of aerosols. No cases of human brucellosis were recorded at this abattoir during the summer of 1980-81.

Murray MJ, Murray AB, Murray CJ. An ecological interdependence of diet and disease? A study of infection in one tribe consuming two different diets. The American journal of clinical nutrition. 1980 Mar; 33(3); 697-701. [PubMed: 7355850].

Abstract: The nature and incidence of infections were studied in two groups of Turkana living in the same area but eating different diets; one consumed milk only and the other a combination of fish and milk. The only apparent and significant nutritional difference between the two groups was mild iron deficiency in the milk drinkers. Episodes of fever, symptomatic infection with malaria and brucellosis, molluscum contagiosum and common warts, episodes of diarrhea, and serological evidence of infection with Entamoeba histolytica were significantly increased in Turkana eating fish. We suggest that this phenomenon may result from a disruption of a long-standing ecological compromise between the all-milk diet of the Turkana and the pathogenic organisms.

Abstract: 418 workers were surveyed at a meat-packing plant. The presence of complement-fixing antibodies to the causative agent of enzootic abortion of sheep (EAS) was detected in 59 workers (14.27%), brucellosis in 32 workers (7.64%) and Q fever in 5 workers (1.18%). EAS antibodies were found to reach titers of 1 : 10 to 1 : 40 and higher, changing dynamically. The persons found to be seropositive belonged mainly to the workers of the sausage-making, slaughtering, intestinal, subproduct and skin-salting departments, as well as to the workers of the sanitary slaughter-house. Such diseases as acute respiratory infections, pneumonia, arthralgia, arthritis, and in women inflammatory urogenital infections, as well as spontaneous abortions were more frequent among the workers seropositive to the causative agent of EAS than in the control groups. Enzootic halprovial abortion of sheep was suggested to be potentially dangerous for certain groups of workers at the meat-packing plant, but the results of the survey indicate that further research in the fields of the epidemiology, clinical picture and laboratory diagnostics of infectious pathology in persons having contacts with the animals infected with the causative agent of EAS is necessary.

Abstract: Data were collected from approximately 1000 dairy herds, initially blood tested for brucellosis in 1977, in each of southeastern and southwestern Ontario. These data were used to evaluate three brucellosis screening programs, the area recertification program, the market cow program and the milk test program. The milk test program was the most efficient program at detecting brucellosis, 29.5% of the herds tested were classified as infected, but lacked the ability to detect a large proportion of "infected" herds (relative sensitivity = 24%). The market cow program was more efficient than the area recertification program at finding infected herds, 3.9% of the herds tested under the market cow program were infected, but had a low relative sensitivity of 12%. The area recertification program was least efficient, 2.3% of herds tested under the area recertification program were infected, but had the highest relative sensitivity (53%). The relative efficiency (predictive value) of the programs was not affected significantly by location of the herds, season of the initial test or herd size. The relative sensitivity of the milk test program was significantly higher in eastern than western Ontario and tended to decrease as herd size increased. The market cow program tended to be more sensitive in the summer months. The relative specificity of the milk test program (0.997) was higher than that of the market cow program (0.960) and the area recertification program (0.884).

Abstract: An epidemiological study of Q fever infection in a Brisbane meatworks was undertaken. A retrospective 10-year study (1968 to 1977) revealed an average annual incidence of 1% for the nine years excluding 1969, while an outbreak with a 7.9% incidence occurred in 1969. No seasonal distribution was found. Results indicated that cattle form the major source of Coxiella burneti (infectious agent of Q fever) in the meatworks and that the highest risk of infection exists on the cattle slaughter floor. The 1969 outbreak showed a departure from the average pattern of infection in that the highest incidences were recorded in small stock (sheep and swine) workers and non-meat handlers rather than in cattle slaughtermen. This suggested the importance of sheep and/or swine forming a source of the infection. Investigations into the factors contributing to the 1969 outbreak in this meatworks failed to reveal an; conclusive data. Inhalation of infectious dust and aerosols proved to be the major channel of transmission of infection rather than direct inoculation. A serological survey of 139 employees revealed 15.8% prevalence of complement-fixing antibodies to C. burneti, 0% to brucellosis, 0.7% to leptospirosis, and 0.7% to Toxoplasma gondii. Of those with positive titres for Q fever, nine of the 22 subjects did not recall any clinical illness.

Abstract: In 1979, 110 people associated with a local rural abattoir presented with an acute febrile illness thought to be Q fever. Of these, 70 were shown by serotesting to have had Q fever, and one was shown to have had leptospirosis alone. Four individuals had mixed infections of Q fever with another zoonotic infection, two with leptospirosis, and two with brucellosis. Only 44% of suspected cases of Q fever were shown to have complement-fixing antibodies to Q fever four weeks after the infection, but 74% had anti-bodies 12 weeks after infection. This epidemic of Q fever occurred soon after the abattoir began to slaughter feral goats for the first time; there is reason to believe that the epidemic may have been related to the introduction of this practice.

Abstract: The importance of disease and management factors in relation to the productivity of traditionally managed White Fulani cattle herds on the Jos plateau is described. Disease per se was not considered to be a major factor limiting productivity with the reduction in importance of the major epizootic diseases of rinderpest and contagious bovine pleuropneumonia. Diseases which were considered of some importance were streptothricosis and liver fluke in adult cattle and coccidiosis and possibly parasitic gastro-enteritis in younger animals. One of the management factors limiting productivity was the keeping of older castrates mostly in the Fulani-owned herds. The most important management practice affecting the productivity of the herds was the provision of dry season supplementation. Only one herd received substantial amounts and its productivity was much greater than that of the other study herds.

Abstract: Seven cases of acute brucellosis seen in Johannesburg during a 9-month period are discussed. Five of these patients probably contracted their infection at the by-products plant of the new abattoir. A survey of culture isolates and serological studies shows a low incidence of acute brucellosis in Johannesburg. The persistently positive serological tests 18 - 24 months after successful therapy emphasize that caution is necessary when interpreting the significance of positive findings. Successful therapy with tetracyline is largely dependent on dosage and length of treatment.

Abstract: Brucellosis is an anthropozoonosis, transmissable between animals and from animals to man. There is no interhuman transmission. Practically all domestic and wild animals are sensitive to various species of Brucella which explains the world-wide occurrence of the disease. Animals transmit the disease to man by direct contact (giving rise most often to professional disease which accounts for approximately 2/3 of the cases in France), and by way of food consumption (milk, cheese). The discovery of host animals may guide the diagnosis of human cases but more often the diagnosis of a human case reveals the existence of a host animal. Among the serological tests which help to follow the course of the human disease, the importance of the card test and of the immunofluorescence reaction are especially emphasized. The incidence of the human disease is difficult to determine: from the number of cases notified, the number of cases actually diagnosed can be estimated (from 3 to 5 times greater in France); however the true number of cases is very difficult to estimate owing to the large number of asymptomatic or atypical cases.

Abstract: Growth of Helicobacter pylori in liquid culture requires the addition of media supplements that often interfere with subsequent purification of bacterial antigens. In order to determine whether cyclodextrins can substitute for conventional H. pylori growth supplements, we cultured H. pylori in the presence of five commercially available cyclodextrins. The effect of these compounds on the production of the vacuolating cytotoxin antigen was evaluated. Several cyclodextrins supported flourishing growth and permitted the consistent production of vacuolating cytotoxin. These data suggest that Brucella broth supplemented with cyclodextrins is an improved medium for bacterial culture and industrial production of H. pylori antigens.

Abstract: Features and protective activity of anti-Brucella murine monoclonal antibodies (Mabs) versus B. melitensis 16M were studied. All Mabs tested were able to confer a good passive protection and showed correlative biological properties in immunological reactions.

Abstract: Two murine monoclonal antibodies, an IgG1 isotype specific for the heavy chain of bovine IgG1, the second an IgG3 isotype and specific for an epitope of the O-polysaccharide of Brucella abortus were conjugated with horseradish peroxidase. The conjugated antibodies were freeze dried in the presence of a number of additives to preserve activity and tested for stability over an 18 month period. Addition of 0.3M trehalose or 0.8% lactalbumin and 3.2% sucrose resulted in the lowest loss of activity if the conjugated antibodies were freeze dried in glass vials. Freeze drying in polypropylene vials resulted in a more rapid rate of deterioration with some additives or an accelerated rate of decline after an initial plateau of lesser loss of activity. The use of polypropylene vials and lactalbumin and sucrose were included in this study because of their low cost compared to glass vials and trehalose. While each antibody behaved differently, addition of trehalose or lactalbumin and sucrose to the conjugated antibodies aided in the preservation of enzyme activity. Both additives provide a suitable method for long term storage of freeze dried or freeze dried and reconstituted monoclonal antibody-enzyme conjugates.

Abstract: Liposomes of stable multilamellar type, which previously demonstrated great efficiency in antibiotic transport, were used in this study as transport vehicles of antigenic extracts of Brucella melitensis (HS: complex of lipopolysaccharide/phospholipids/outer membrane proteins). The incorporation of HS into positively charged liposomes produced a protective effect against experimental murine brucellosis when they were administered 1 day before or 2 days after infection, as the number of colony-forming units in the spleen was reduced in relation to the untreated control group (P < 0.01). On the other hand, the use of HS-free or bound in liposomes with negative net charge did not produce a significant effect. Moreover, the incorporation of HS into cationic liposomes eliminated the toxicity of the lipopolysaccharide.

Abstract: In 1992, the North Carolina Department of Environment, Health, and Natural Resources received 18 case reports of brucellosis from a county health department. All patients had potential exposure to the kill floor of one pork processing plant. A subsequent National Institute for Occupational Safety and Health health hazard evaluation surveyed 154 (99%) of 156 kill floor workers of this plant and found that 30 (19%) had evidence of recent (or persistent) brucellosis. These data show that significant exposure to Brucella is occurring among packing plant workers in North Carolina and suggest that some of the approximately 38,000 production workers in pork processing plants in the United States are at risk of contracting swine brucellosis. Additional measures may need to be taken to prevent occupational exposure to Brucella.

Abstract: The capacity of liposomes constituted by dycetyl-phosphate (0.009 mM), cholesterol (0.017 mM), lecithin (0.003 mM), and myristic (0.1 mM), stearic (0.1 mM), or oleic acid (0.1 mM) to modify the lymphocyte response to Brucella melitensis antigens in mice was studied. Mice treated with antigens mixed with liposomes containing myristic, stearic or oleic acid had higher antibody titres than mice given antigen suspended in a saline solution. Liposomes alone, without Brucella antigens, resulted in increased 3H-thymidine incorporation by lymphocytes both in vivo and in vitro. The addition of polyclonal activators (LPS and ConA) caused a further increase of 3H-thymidine uptake. Moreover, spleen lymphocytes from mice inoculated with Brucella antigens mixed with the liposomes had a significantly lower population of B lymphocytes (10%), and a notable increase in the Tc lymphocytes (20%). Autoradiography of sections of popliteal ganglia of treated mice showed that the radioactivity was concentrated mainly in the membrane structures of the cell.

Abstract: Human brucellosis, a multisystem disease which may mimic other conditions, has a low incidence in childhood and the diagnosis may easily be missed. Over a 7-month period 9 children with brucellosis presented to the Department of Paediatrics and Child Health, Tygerberg Hospital. Six of the children had consumed unpasteurized milk. The main presenting symptoms were fever, fatigue, headache, myalgia and haematuria. Clinical signs included lymphadenopathy (3), nasopharyngitis (2), features of lower respiratory tract infection (2), splenomegaly (2) and pyrexia (1). The diagnosis was made on the basis of a positive serological titre (> 1:160) for Brucella abortus. The prozone phenomenon was encountered in 6 cases; however, the Coombs test confirmed the diagnosis in these cases. Children under 7 years were treated with co-trimoxazole and rifampicin and those over 7 years with tetracycline and rifampicin, for at least 6 weeks. No relapses were detected on follow-up.

Abstract: As the result of the complex clinico-laboratory examination of 265 cattle breeders working on 5 farms where cases of brucellosis among agricultural animals had been detected, 62 persons (21.8%) were found to have the positive reaction to brucellosis. A strikingly high proportion (38.3%) of infected persons was found among the workers of a cattle-feeding complex. Out of 62 persons with the positive reaction to brucellosis, 32 had different clinical manifestations of the disease. Treatment for all persons who needed it was organized, the rest were placed under dispensary observation. Regular complex clinico-laboratory examinations of cattle breeders to brucellosis were recommended.

Abstract: Between July 1991 and August 1992, 63 Jordanian dairy farms selected by stratified random sample were visited to identify the major causes and prevalence of intramammary infections in dairy cows. Of 773 cows examined 60% of all sampled quarters had > 283,000 cells/ml. The mean value of somatic cell count (SCC) was positively associated with age in lactations and negatively with herd size. Cows milked by bucket milking machines or in fully automatic parlours had a lower mean SCC than those milked by hand. Many management faults pertaining to milking procedure and maintenance of milking machines were noted. The most common isolate from clinical cases was Staphylococcus aureus (37.5%). Estimates of prevalence of bacterial pathogens in intramammary infections were: coagulase negative staphylococci (16.04%), S. aureus (9.41%), Klebsiella spp. (6.17%), Corynebacterium bovis (5.35%) and Brucella melitensis (4.52%). The results demonstrate the essential need for the development of a national mastitis control programme.

Abstract: The introduction of U H T treatment of milk into South Africa raises problems for local public health authorities controlling milk hygiene in their respective areas because of the legislation concerning milk produced and sold for various purposes. It is essential that the raw milk supply be derived from healthy cows with healthy udders, and be produced under sanitary conditions. Uniform control standards are necessary for U.H.T. milk and other forms of milk supplying throughout the Republic. Uniformity is also desirable for efficient local authority veterinary health services. The progress of veterinary control and supervision of Johannesburg's milk supply is discussed to indicate what has been achieved. There is need for a national scheme for controlling mastitis. Tests for the purchase of milk on quality must include tests for udder health by regular electronic cell counting of raw milk sources. Appropriate mastitis control incentive or penalty schemes for milk producers should be instituted.

Abstract: We have studied two isoforms of Nef, Nef-27 and Nef-25, which were produced in E. coli. Nef-25 lacked the first 18 N-terminal residues of Nef-27 and both were nonmyristylated. Nef-27 fuses small unilamellar dipalmitoyl phosphatidylcholine vesicles (SUVs), as indicated by enhanced light scattering of SUVs and lipid mixing using concentration-dependent fluorescence dequenching. Nef-27 also causes the appearance of a shifted isotropic peak in the 31P NMR spectra of these vesicles, suggesting that protein interactions induce nonlamellar lipid structures. Recombinant Nef-25, which lacks only the 18 N-terminal residues of Nef-27, does not fuse vesicles and has little effect on the 31P NMR spectra. On the other hand, synthetic peptides consisting of 18 or 21 of the N-terminal residues of Nef-27 are strongly membrane perturbing, causing vesicle fusion and inducing isotropic peaks in the 31P NMR spectrum. Endogenous fluorescence spectra of the N-terminal peptide (21 residues) with SUVs show that the N-terminal sequence of Nef may achieve these perturbing effects by inserting its hydrophobic side into the lipid bilayer. Theoretical calculations using hydrophobic moment plot analysis indicate that short-length stretches (i.e., six amino acid residues) of the N-terminal sequence may insert into the lipid bilayer as multimeric alpha helices or beta sheets. The above-described membrane activities of Nef-27, which principally reside in its N-terminal domain, may play critical role(s) in certain functional properties of the full-length protein. For example, the fusogenic activity of the N-terminal sequence may be involved in the extracellular release of Nef-27, much of which appears to be associated with small membrane vesicles. The fusion activity may also be relevant to the ability of Nef-27 to downregulate CD4 and IL-2 receptors when this protein is electroporated into cultured lymphocytes, an activity not possessed by Nef-25.

Abstract: Different macrophage subsets are present in the spleen, i.e., marginal zone macrophages (MZM), marginal metallophilic macrophages (MMM) and red pulp macrophages (RPM) and all are depleted by a single treatment with liposome-encapsulated clodronate. These macrophages can be distinguished by differences in localisation patterns, membrane antigens and repopulation kinetics after depletion. In experiments on the involvement of splenic macrophages in the humoral immune response, there was an acceleration of the repopulation kinetics of all macrophage subsets in the spleen after intravenous injection of an autoclaved suspension of Brucella abortus 544 (BA 544 antigen). The time required to obtain 100% repopulation in macrophage-depleted control mice was 2 weeks for RPM, 6 weeks for MMM and > 2 months for MZM. However, after BA 544 injection, 100% repopulation was obtained within 4 days in the case of RPM and within 2 weeks in the case of MMM. Acid phosphatase activity, indicating the presence of MZM, had returned to normal levels within 2 months. Acceleration of repopulation was observed only after intravenous administration of antigen preparations from Brucella strains (except strain BA 19). Although BA 544 antigen stimulated the proliferation of precursors of all of the macrophage subsets in the spleen and liver, it also affected mature members of the mononuclear phagocyte system such as MZM and dendritic cells in the spleen.

Abstract: From 1973 through 1992, 426 cases of human brucellosis were reported in California, of which 98% were laboratory confirmed. Brucella melitensis was identified in 185 cases (78.7% of the bacteriologically typed cases). Hispanics accounted for 81% of the cases from 1983 to 1992 compared with 65% during the previous decade (P < .01). The population-adjusted average annual incidence was higher in Hispanics, especially in children and teenagers, compared with non-Hispanic whites and African Americans. Slaughterhouse cases decreased from 25% during 1973-1982 to < 3% during the following decade. Changes in case distribution were characterized by a decreasing incidence in the Central Valley and an increasing incidence in the San Francisco Bay area and the southern Coast Range. Hispanics were more likely to report being infected by consumption of milk and cheese in Mexico during 1983-1992 than during the previous 10 years (relative risk, 1.45). Between 1973 and 1992, human brucellosis in California evolved from an occupational to a foodborne illness.

Abstract: The diagnostic value of bacteriological and serological methods for the laboratory diagnosis of brucellosis was studied. In the analysis of milk and cheese specimens Brucella cultures were isolated and differentiated as B.melitensis, biovar I, and B.abortus, biovar 4. In 25.6% of cases B.melitensis culture, biovar 1, was isolated from the blood of persons suspected for brucellosis. The isolation of B.melitensis culture from milk showed that this infective agent migrated from small animals to cattle, which was indicative of a high risk of human infection in the state of Zacatecas, Mexico. The comparative evaluation of serological diagnostic methods (the agglutination test in test tubes, Huddleson's slide test, the acidic rose bengal test and the 2-mercaptoethanol test) showed high sensitivity of rapid tests (Huddleson's test and the rose bengal test in 93.7% and 87.9% of cases respectively). The 2-mercaptoethanol test which gave positive results in 63.8% of cases provided additional information characterizing the course of infections process.

Abstract: Brucellosis is an infectious zoonosis. It is a occupational disease of stock-breeders, workers of slaughter houses and veterinarians. Brucellosis is characterized by a variety of clinical symptoms and involves several organs of the body. Skeletal manifestations often facilitate diagnosis. Difficulties in diagnosis of brucellosis in Poland are resulted from an increased amount of low-symptomatic or asymptomatic cases, a decrease in acute brucellosis and mild course of chronic disease. Therapy is related to the stage of the disease, antibiotics are required in acute brucellosis. Symptomatic treatment with special emphasis to neurological and psychiatric management is used in patients with chronic brucellosis. Early made diagnosis is important because undiagnosed disease leads to severe disability and loss of work ability.

Abstract: Serologic test data and bacteriologic culture results from 816 cattle were evaluated. Brucella abortus (field strain or strain 19) was isolated from 27.3% of the cows. Results of the card test, particle concentration fluorescence immunoassay (PCFIA), and complement fixation (CF) test were compared. Antibody titers were directly associated with B. abortus isolation, with the majority (64.7%) of field strain isolations having a PCFIA value of < or = 0.300 and a CF test result of at least 1+ at a 1:160 dilution. The specificity of the CF test was significantly higher than that of the PCFIA (at CF > or = 1:40 and PCFIA < or = 0.300).

Abstract: Brucellosis has always been an unusual disease in children and, concomitant with the control of the disease in domestic animals, reports have become sparse. The pediatrician, therefore, may not be aware of the protean clinical manifestations of childhood brucellosis. In 1973, nine cases occurred during a three-month period in El Paso, Texas. All cases were marked by spiking fevers and lethargy of four days to four weeks in duration. Tender hepatomegaly or splenomegaly was striking in seven patients. Other characteristics included epistaxis, arthralgia, myalgia, and weight loss. Leukopenia and leukemoid reaction were found in five patients. All of the patients tested had elevated liver enzymes. Febrile agglutinins were invaluable in screening for an early clue to diagnosis. When Brucella abortus antigen agglutinated serum from patients with a positive screen in dilutions greater than 1:320, a presumptive diagnosis of brucellosis was made. Brucella was isolated from the blood or bone marrow in seven patients and the time of incubation proved crucial for successful recovery. Bacterial blood cultures are usually discarded at ten days of age, as were cultures from the only two patients from whom the organism was not recovered. All of the cultures incubated for 12 to 15 days grew B. melitensis, an unusual causative species in the United States. However, several patients admitted eating cheese from the State of Chihuahua, Mexico, made from unpasteurized goat's milk, the presumed source of the infection. Within one to three days, all patients responded dramatically to antibiotics; tetracycline was given orally for 21 days and streptomycin intramuscularly for 14 days. Pediatricians caring for patients in areas where consumption of unpasteurized milk products is likely would do well to consider brucellosis in a child with obscure fever or toxic hepatosplenomegaly.

Abstract: The use of a fluid-enrichment technique increased by 10-16% the recovery rate of Brucella abortus from milks of herds positive to a milk ring test. Three solid media for direct cultural techniques were also tested on the same milks. Combinations of these techniques produced a recovery rate of ca. 88% from MRT-positive-herd milks using a mixture of the centrifuged deposit and cream layer as the inoculum as against a recovery rate of ca. 75% using the MRT cream layer as inoculum. B. abortus biotypes 1-5 and 9 were isolated and growth of all the biotypes was supported by all media used.

Abstract: Five calves from Brucella abortus reactor cows bought after they had been on infected farms for two to six weeks, were reared in isolation and examined serologically for approximately six months prior to cultural and biological examination post mertem. One calf which had been serologically negative to the Rose-Bengal, serum agglutination and complement fixation tests for the last eight weeks of its life yielded a culture of Br abortus biotype 1. This serotype had also been isolated from its dam at parturition.

Abstract: An outbreak of Brucella melitensis in a family was studied. From the fourteen family members who ate unpasteurized goat cheese nine became ill. Patients included four females and five males of 8 to 75 years old. In seven of the patients the diagnosis was confirmed by positive blood culture for B. melitensis biovar 1. All the patients were analyzed by standard tube agglutination (STA) and standard tube agglutination with 2-mercaptoethanol (STA-2ME) tests at the time of diagnosis. In six of the patients, ELISA assays were used to assess the humoral immune anti-protein and anti-lipopolysaccharide (LPS) responses. Anti-LPS IgG antibodies were detected in all of the patients. Anti-proteins IgG antibodies were present at significant levels in all the studied patients including the STA-2ME negative ones.

Abstract: Two types of superoxide dismutase (SOD) have been found in Brucella abortus, a cytosolic Mn-SOD and a Cu/Zn-SOD of unknown location. We sought to determine the subcellular location of Cu/Zn-SOD in B. abortus ST 19. We report a modified spheroplasting procedure for the release of periplasmic contents from B. abortus cells using a dipolar ionic detergent, Zwittergent 316. This detergent, used in place of EDTA, destabilizes the outer membrane sufficiently to allow penetration of lysozyme and the subsequent selective release of periplasmic proteins by osmotic shock. Cytoplasmic cross-contamination of periplasmic fractions was assessed by assaying for malate dehydrogenase activity. Cyanide-sensitive and cyanide-insensitive SOD activity was measured by both the xanthine oxidase-cytochrome c method and a hematoxylin assay. Results suggest that B. abortus Cu/Zn-SOD activity is periplasmic. This zwittergent-lysozyme extraction procedure may be applicable to the separation, isolation and characterization of many other periplasmic proteins of B. abortus and other Gram-negative organisms especially when cytosolic contamination is undesirable.

Abstract: Performing the milk ring test in herd samples from herds of approximately twenty dairy cows is an acceptable method in checking herds for brucellosis. Herds of approximately forty dairy cows or more are unsuited for this method of checking. For the above reasons, it is essential that the compulsory examination of the blood of aborting cows should be consistently carried out and that the larger herds in the threatened areas should be examined by performing the milk ring test in individual samples at regular intervals.

Abstract: Defatting milk for the Brucella abortus whey agglutination test with chloroform caused some reduction in the titre in i.u./ml. in 42 of 57 samples and this reduction exceeded 50% in seven. Though this procedure will probably rarely affect the final interpretation of the test defatting the milk by centrifugation is preferred.

. From the Centers for Disease Control and Prevention. Brucellosis outbreak at a pork processing plant--North Carolina, 1992. JAMA : the journal of the American Medical Association. 1994 Jun 8; 271(22); 1734-5. [PubMed: 8196106].

Abstract: Over 55% of 7161 people examined in different parts of Western Nigeria have positive Brucella abortus-antibodies in their sera. Higher incidence of titres were found among dairy farmers and slaughtermen than in the general population. The rates of infection among the human and the cattle populations in two farms studied were very similar. There was evidence of high rate of cross-infections going on in one of these farms: more than 63% of the herd in this farm showed serological evidence of active brucellosis. The socio-economic implications of high incidence of bovine brucellosis in a rapidly developing country, like Nigeria, are outlined. The need for the establishment of public health laboratories to facilitate early diagnosis of active disease and the necessity for full collaboration between the medical and veterinary professions for the proper control (and eradication) of brucellosis in Nigeria are discussed.

Abstract: The prevalence of brucellosis among abattoir workers in Saudi Arabia was determined through a randomised multi-stage sampling of 1200 abattoir workers. Diagnosis was made by both blood culture and standard tube agglutination test. The overall prevalence of brucellosis was 4.0% among abattoir workers. Infection was more common among butchers (8.9%), veterinarians and veterinary assistants (5.4%), and administrative personnel (1.1%). In order to reach optimum planning for a national brucellosis control programme in Saudi Arabia, ministerial co-ordination should be established with formulation of inter-governmental veterinary agreements between the Saudi government and the relevant authorities of animal exporting countries. A national survey on animal marketing is required.

Abstract: In Experiment 1, polychlorinated biphenyls (Aroclors 1016, 1232, 1242, 1248, and 1254) were fed ad libitum to White Leghorn chickens at dietary levels of 5, 10, and 20 p.p.m. for 8 weeks. Hatchability of eggs from hens fed 10 and 20 p.p.m. of Aroclors 1232, 1242, and 1248 was reduced. The weights of the spleen and bursa of Fabricius of day-old chicks from hens fed 10 p.p.m. of Aroclors 1242 and 1248 or 20 p.p.m. of Aroclors 1232, 1242, 1248, and 1254 were less than those of control chicks. Body weights were similar to those of controls at hatching, but the 3 week weight gains of progeny from hens fed 20 p.p.m. Aroclors 1242 and 1248 were less than gains of control progeny. After replacement of the PCB diet with uncontaminated feed, hatchability improved within 2 weeks and by 8 weeks was similar to the hens were fed uncontaminated feed. In Experiment 2, hens were fed 10 p.p.m. 1248 ad libitum for 8 weeks. The progeny bursa weights were reduced, but the treatment did not interfere with interfere with antibody production. PCB treatment had no effect on antibody production after primary or secondary challenges with Brucella abortus. However, after the primary challenge, bursectomized birds of both the control and PCB groups produced significantly fewer antibodies than the nonbursectomized birds. Differential accumulations of the polychlorinated biphenyls occurred in the effs. Chlorinated biphenyls with relative retention times less than that of DDE accumulated to less than one-half of the dietary concentration whereas chlorinated biphenyls with retention times greater than that of DDE accumulated in the eff to a concentration equivalent to the dietary concentration.

Vassallo DJ. The centenary of the sinking of the Mediterranean Fleet flagship, HMS Victoria. What was the role of Malta fever?. Journal of the Royal Naval Medical Service. 1993 Summer; 79(2); 91-9. [PubMed: 8263860].

Abstract: This article commemorates the centenary of the tragic loss of the battleship HMS Victoria and 358 of her crew, together with the Commander-in-Chief of the Mediterranean Fleet, Vice-Admiral Sir George Tryon, as a result of an enigmatic order by Tryon himself. It also investigates the medical aspects and explores the contention that Tryon was suffering from Malta or Mediterranean fever (Brucellosis).

Abstract: Forty-five brucella infected cows were studied 15 to 30 days following abortion. Eighteen cows were slaughtered and autopsied for examination of Brucella in the organs and lymph nodes. The remaining 27 cows were treated during two and one-half months and then slaughtered after undergoing a treatment with 0, 1, 2 or 3 intraperitoneal injections of oxytetracycline dissolved in 100 ml of physiological saline. The treatment modified only slightly the natural evolution of antibody titers. The levels of infection were similar for all cows which received no treatment. Cows treated with oxytetracycline had less severe infection than the non treated animals and four were infection-free at slaughter. The level of infection of treated cows was independant of the treatment regime. The advantages of treating non pregnant cows to reduce the level of infection and risk of abortion were discussed.

Abstract: The detection of Brucella bacteremia by subculture does not always correlate with a positive signal in the BACTEC NR730 nonradiometric system (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.). The effect of the inoculum size, pH, sodium polyanetholesulfonate, carbon sources (i-erythritol, sodium pyruvate, monosodium glutamate, D-glucose, and L-alanine), and urea in the release of CO2 was evaluated by using the reference strain Brucella melitensis 16M. In standard NR6 vials with or without blood, inocula 5 to 10 times larger (at least 265 CFU per vial) than those usually found in the blood of patients with brucellosis were necessary to produce a positive growth value (GV) in 4 days or less, and similar results were obtained with vials supplemented with the substrates listed above. GVs were consistently lower in vials with sodium polyanetholesulfonate than in vials without this agent. Vials with no blood inoculated with 265 CFU per vial showed turbidity 1 day before GVs became positive, proving that the major limiting detection factor was the low level of release of CO2 and not an inadequate growth medium. In NR6 vials buffered to pH 6.2, GVs became positive faster and were higher than those in standard vials. NR6 vials at pH 6.2 with 0.3% sodium pyruvate yielded a positive GV in the first day of bacterial turbidity.

Abstract: A risk assessment model is presented, for use by local decision-makers to aid the evaluation of proposed changes in existing brucellosis eradication or control programmes. This model provides a format and structure for gathering and analysing data. The model uses data which are generally available and accessible, so that minimum time, expense and effort are required for collection. The use of this model enables an estimation of the risk of introduction of brucellosis into a non-infected population, based on the probability of importing the agent and subsequent spread, given the existence of specified surveillance and control measures. The model creates a point estimate of the risk associated with a given set of conditions.

Abstract: This paper presents a quantitative risk assessment method based on the portrayal of risks using scenario trees, the translation of evidence into probability curves and the aggregation of scenarios using Latin Hypercube simulation. An example of a quantitative risk assessment on the importation of swine for slaughter illustrates the interpretation of evidence of two swine disease risks: pseudorabies (Aujeszky's disease) and brucellosis.

Abstract: Bacteriologic culture of udder secretions and card, rivanol, and complement fixation tests for detection of brucellosis were performed on samples from 828 cows vaccinated with strain 19 in herds with brucellosis. An ELISA and a particle concentration fluorescence immunoassay were performed on sera from 560 and 569 cows, respectively. A field strain of Brucella abortus or strain 19 was isolated from 278 cows. The ELISA and the card test had high sensitivity, but low specificity. Data suggested that there is no advantage to using primary binding assays rather than the simple buffered antigen agglutination procedures to detect cows infected with a field strain of B abortus.

Abstract: Several particulate antigens and liposomes were intravenously injected in mice in order to study their localization patterns in spleen and liver. Liposomes have been proposed as promising carriers for haptens and antigens. It was studied whether the phospholipid composition, cholesterol content and charge of the liposomes played a role in their distribution within the spleen. Different thymus-independent type 1 and type 2 and thymus-dependent particulate antigens as well as liposomes were labeled with the lipophilic fluorochrome Di-I. After labeling they were intravenously injected and spleens and livers were removed at different time intervals and prepared for light- and fluorescence-microscopy. We have observed that all particulate antigens and liposomes administered to the mice localized according to the same distribution pattern in the spleen. After 2 and 4 h particles were located in macrophages of the marginal zone and after 24 h white pulp macrophages had also ingested particulate antigens and liposomes. So we conclude that the distribution of the particulate antigens and liposomes in the spleen is independent of the immunological nature of the particles. Results are discussed with respect to the question whether or not the distribution of particulate antigens and liposome associated antigens or haptens, may be a crucial factor in determining the type of immune response to be elicited.

Abstract: The sample studied contained 214 people divided into 2 groups: 107 abattoir workers and 107 blood donors. All the sera were analyzed by testing with buffered antigen. The percentage of positive sera in the exposed population was 14% whereas it was zero in the group of blood donors.

Abstract: We describe our experience with 73 patients diagnosed with brucellosis during the years 1979-91 at two Jerusalem hospitals: Hadassah Mount Scopus (37 patients from 1979-1984) and Shaare Zedek (36 patients from 1979-1991). The patients included 32 children less than 14 years old and 41 adults; 70 of the patients were non-Jews. In all cases the pathogen was Brucella melitensis. The high proportion of children and the equal sex distribution was quite different from the age and sex distribution of brucellosis in Western countries where it is more common in adult males, and similar to that reported from other near-Eastern countries where household dairy products, and not occupational exposure, are the most common source of infection. The short duration of disease (< 2 weeks) prior to diagnosis in 70% of the patients is attributed to the ready availability of appropriate medical care, and a very high index of suspicion for brucellosis in the Jerusalem non-Jewish population. Abdominal symptoms were more common in adults, whereas enlarged lymph nodes and liver, skin rash and pharyngitis were more frequently observed in children. Some of these differences may be attributed to the very short duration of disease in most children at the time of presentation. Combination therapy with tetracycline-streptomycin or tetracycline-rifampin yielded superior results as compared with single-drug treatment in terms of early defervescence and relapse rates. The present experience underlines the importance of endemic brucellosis which still represents a significant public health problem in children and adults in Mediterranean countries.

Abstract: 1. Beta-adrenergic agonist (Clenbuterol, 0.33 mg/kg) and corticosterone (10 mg/kg) were incorporated into a diet based on maize and soybean meal. Their effects on performance, carcase composition, hepatic microsomal mixed function oxidase and antibody production were investigated in female broilers. 2. Dietary corticosterone reduced the titre to sheep red blood cells, while it was unchanged by clenbuterol. 3. Clenbuterol exerted a promoting effect on gain-to-food ratio, carcase protein and hepatic microsomal cytochrome P-450 content. 4. Addition of clenbuterol to the corticosterone-containing diet prevented the increase in abdominal fat weight and uric acid excretion induced by corticosterone, but did not affect total fat mass. 5. The results showed that clenbuterol reduced abdominal rather than carcase fat and prevented protein degradation in the body when chicks were treated with corticosterone. Clenbuterol also influenced the content of cytochrome P-450, but not the humoral immunity.

Greenstone G. Brucellosis: a medical rarity that used to be common in Canada. CMAJ : Canadian Medical Association journal = journal de l'Association medicale canadienne. 1993 May 1; 148(9); 1612-3. [PubMed: 8477393].

Abstract: Because the Australian livestock industries rely overwhelmingly on year-round grazing of pastures, nematode parasites are of major concern. Nematode parasitism is recognized as the cause of the most economically important diseases of sheep, while in cattle, the relative importance of nematode parasitism will probably increase following the eradication of diseases such as brucellosis and tuberculosis. The extraordinarily high prevalence of resistance to benzimidazoles, levamisole and morantel in nematodes parasitising sheep has stimulated the formulation and, to a surprisingly large extent, adoption by farmers of strategic control programmes aimed at preserving the useful life of ivermectin. Even so, it is highly likely that we will enter the 21st century with ivermectin resistance. Current research aimed at control in the next century is progressing in three broad directions. First, towards better control using existing anthelmintics through modifications to pharmacokinetics, together with integrated management aided by computer simulation models of nematode population biology. Secondly, towards immunological control arising from a better understanding of the ruminant immune response to helminths, particularly in young animals, and finally, a coordinated investigation of genetic resistance, within breeds, of sheep to nematode parasitism. The appearance in the 21st century of novel anthelmintic compounds, most probably as a result of research by the international pharmaceutical industry, may well reduce the sense of urgency driving current Australian research. Nevertheless, success in one or more of these three lines of endeavour will mean a longer and more profitable life for the next generation of anthelmintics marketed in Australia.

Abstract: Contemporary development of agricultural structures in connection with changes of socio-economic relationships complicates the task of health care for rural population. Financial difficulties disordered managing and control of ecologic situation in rural area. The causal matters include absence of new agricultural machinery, chaotic use of agrochemicals purchased abroad, increased occurrence of occupational brucellosis and other diseases in animal husbandry farms. The authors necessitate effective control provided by industrial medical officers and regional sanitary and epidemiologic service to improve work conditions and lower morbidity and traumatism in rural territories.

Abstract: Rheumatological manifestations are frequently reported in patients with brucellosis. In a retrospective study of 90 patients diagnosed with brucellosis over a period of 18 years, 83 (92%) patients were Bedouins, 55 of whom (61%) reported ingestion of unpasteurized goat milk and goat milk products. The male/female ratio was 1:1, and the adult to child ratio was 3:2. The mean age of the patients was 25 years (range, 1-72 years). Rheumatological manifestations (myalgia, arthralgia, and arthritis) were reported in more than half of the patients. These manifestations started on days 3 and 4 of the disease and were mild to moderate in severity. Myalagia was evident in 49 (54%) patients and was more common in adults than in children (67% versus 37%; P < .01) and in men (67%) than in women (42%; P < .01). Arthralgia was the most common musculoskeletal manifestation, found in 55 (61%) patients, and occurred more often in children than in adults (74% versus 52%; P < .05). Arthritis was detected in 37 (41%) patients. The hip and knee joints were the most common sites of arthritis (31% each) followed by sacroiliac involvement (17%) and shoulder or spine involvement (5% each). Arthritis was also more common in children (63% versus 29%; P < .01). The prevalence of arthritis was similar in men and women. Cure was achieved in all patients after antibiotic therapy.

Abstract: In patients with polytrauma or major surgery, severe bacterial infections leading to septic shock and multiorgan failure are still a major cause of death. Prevention of septic shock in patients at risk would be an alternative to treatment of patients with overt septic shock. We therefore conducted a trial with the monocyte activator muramyl tripeptide phosphatidylethanolamine (MTP-PE) in an experimental pig model. Liposome encapsulated MTP-PE (50 micrograms/kg of body weight) or liposomes alone were given intravenously at 72 or 24 h before endotoxemia was induced by lipopolysaccharide (LPS), simultaneously with the induction of endotoxin shock, or 1 h thereafter. Pretreatment with MTP-PE at 72 and 24 h before endotoxemia was induced resulted in a reduction of endotoxin shock-induced mortality from 81.8% (9 of 11 animals) in the control group to 8.3% (1 of 12 animals) of the MTP-PE-pretreated animals (P < 0.001). The administration of MTP-PE 24 h before the induction of endotoxin shock was more effective (P < 0.01) than administration of MTP-PE 72 h before endotoxemia was induced (P = 0.05). The pretreated animals did not develop fever or cardiovascular complications, and pulmonary function was significantly improved. Furthermore, the alpha-form of the soluble CD14 LPS receptor in pig serum showed a marked decrease in LPS-treated animals, and this decrease was reduced by MTP-PE pretreatment at 24 h before endotoxemia was induced. When MTP-PE was given simultaneously with the induction of septic shock or 1 h thereafter, it did not influence either mortality or morbidity. In conclusion, pretreatment of pigs with MTP-PE improves several parameters of endotoxin shock and it reduces mortality. Patients with high risk of developing septic complications might benefit from a pretreatment with this monocyte-activating substance.

Abstract: A versatile method for the extraction of Brucella DNA and PCR are presented as reliable tools for the detection of Brucella spp. from body fluids of infected animals. Two oligonucleotides homologous to regions of the gene encoding for an outer membrane protein (omp-2) were designed to detect the pathogen from milk and/or blood of infected goats, bovines, and human patients. The sensitivity of our test and its ability to detect the pathogen in samples from the field reveal a promising advance in the diagnosis of brucellosis in animals and humans.

Abstract: A study was performed to evaluate the previously described PCR (C. Romero, C. Gamazo, M. Pardo, and I. López-Goñi, J. Clin. Microbiol. 33:615-617, 1995) for the diagnosis of brucellosis in dairy cattle. Milk samples from 56 Brucella milk culture-positive cattle and from 37 cattle from Brucella-free herds were examined for Brucella DNA by PCR and for specific antibodies by an indirect enzyme-linked immunosorbent assay (ELISA). The specificities of both tests were 100% when testing the milk samples from Brucella-free cattle. The milk samples from 49 infected cattle were positive by PCR (87.5% sensitivity), and 55 were positive by ELISA (98.2% sensitivity). A PCR-positive sample was negative by ELISA, and 7 ELISA-positive samples were PCR negative, yielding an observed proportion of agreement of 0.91 for the two tests. Although the results suggest that ELISA is a better screening test than PCR, the combined sensitivity of the two assays was 100%, and their simultaneous application could be more useful than one test alone for a rapid screening of brucellosis in dairy cattle.

Abstract: The authors describe an attempt to control Brucella melitensis infection in a large camel herd in Saudi Arabia. Sera from the entire herd (2,536) were examined by the Rose Bengal and standard United States of America buffered plate agglutination tests. The overall Brucella seroprevalence was 8%. Milk samples from the 120 seropositive milking camels were cultured on Brucella-selective media. B. melitensis biovars 1, 2 and 3 were isolated from 41 camels (34%). Seropositive camels (202) were treated for the first time with a combination of long-acting oxytetracycline (OTC) at a dose of 25 mg/kg administered intramuscularly (i.m.) every 2 days for 30 days and streptomycin at 25 mg/kg i.m. every 2 days for 16 days. In addition, milking camels were given OTC-intramammary infusion at a rate of 10 ml/teat every 2 days for 8 days. This regimen was found to be effective in eliminating the shedding of Brucella organisms by camels, with no relapse. Moreover, all treated camels became seronegative within 16 months after treatment. Seronegative camels (2,331) were vaccinated for the first time with the B. melitensis Rev. 1 strain vaccine, as follows: a) 175 young camels (aged three months to one year) were each inoculated subcutaneously with a full dose (1-2 x 10(9) viable organisms in 1 ml). Brucella antibody titres between 1:50 and 1:200 were detected 2-4 weeks post-vaccination. Brucella antibodies decreased gradually until the animals became seronegative 8 months after vaccination. b) 2,156 camels aged more than one year were each inoculated subcutaneously with a reduced dose (1-2 x 10(6) viable organisms in 1 ml). Antibody titres measured 2-4 weeks post-vaccination varied from 1:25 to 1:200. The titres decreased gradually, until the animals became seronegative 3 months post-vaccination. No Brucella organisms were recovered from repeated udder secretion samples from all vaccinated milking camels, and no abortions were recorded among pregnant vaccinated camels.

Abstract: The 16S-23S rRNA spacer regions of Brucella abortus, B. melitensis, and B. suis were cloned and subcloned after PCR amplification. Sequence analysis of the inserts revealed a spacer of about 800 bp with very high ( > 99%) homology among the three species examined. Two genus-specific primer pairs, BRU-P5-BRU-P8 and BRU-P6-BRU-P7, that could be used in a nested PCR format and three genus-specific DNA probes, BRU-ICG2, BRU-ICG3, and BRU-ICG4, were deduced from this spacer. The specificity and sensitivity of both primer sets and probes were examined by testing them against a collection of 18 Brucella strains and 56 strains from other relevant taxa by using PCR and the Line Probe Assay (LiPA), respectively. A method for direct detection of Brucella spp. in 1 ml of raw milk was developed on the basis of enzymatic treatment of the milk components and subsequent PCR and LiPA hybridization. After a single PCR, sensitivities of 2.8 x 10(5) and 2.8 x 10(4) CFU/ml were obtained for detection by agarose gel electrophoresis and LiPA, respectively. Nested PCR yielded a sensitivity of 2.8 x 10(2) CFU/ml for both methods.

Abstract: The possibility of using an ELISA for the diagnosis of ovine brucellosis in milk (M-ELISA) was investigated. The aim of the study was to establish whether the specificity and sensitivity of the M-ELISA would be high enough to detect low levels of Brucella antibodies in ewe milk. The diagnostic performances of the test under study were established by means of reference standards and compared with conventional screening and confirmatory tests under field conditions. The diagnostic specificity of the M-ELISA established on a number of samples from Brucella-free flocks was 100% while relative to RBT and CFT positive reactors the M-ELISA demonstrated sensitivity of 65 and 83% respectively. Its sensitivity relative to culture positive animals was of 92%. The course of Brucella antibodies in milk of positive sheep was evaluated in colostrum and in mature milk for a period of 30 days after delivery and it appeared that concentrations of immunoglobulins in milk tend to sharply decrease soon after parturition while in blood serum these remained constantly high. It was concluded that the M-ELISA for Brucella antibodies in ewe milk can be regarded as a complementary diagnostic tool for individual testing but it would be poorly viable if used as a screening test applied to pooled flock milks.

Abstract: Brucella milk ring tests (BMRT's) were performed on fresh herd milk samples and pooled samples, preserved at a processing factory with potassium dichromate. Preserved milk from samples pooled over a 10-day period, gave results that were similar to an average result for individual fresh daily samples over the same period. When fresh samples gave variable results and the pooled preserved sample gave a negative result, it was considered that the latter result was more reliable, possibly due to factors causing false positive reactions being diluted. Blood testing of herds for circulating antibody indicated that the use of either fresh or preserved milk samples could occasionally produce false negative or false positive results.

Abstract: Fifty-two cases of childhood brucellosis which occurred in north-western Greece during the 15-year period 1979-1993, are reviewed. It is believed that they represent very closely the total incidence of the disease in the region which has a population of 100,000 children aged 0-14 years old. Brucellosis-affected children were almost exclusively from goat- or shepherd families and of both sexes and all age groups. A broad spectrum of clinical manifestations ranging from malaise only to brain abscess was observed. Fever and arthralgia were the most common manifestations followed by malaise, myalgia, sweating, rash, cough, and gastro-intestinal, cardiac and CNS involvement. Splenomegaly was found more often than hepatomegaly and lymphadenopathy. Laboratory findings included anaemia, leukopenia, neutropenia, lymphocytopenia, monocytosis, eosinophilia, thrombocytopenia and pancytopenia. Leukocytosis and lymphocytosis were extremely rare and ESR and serum C-reactive protein levels were mildly elevated. All patients had positive Rose Bengal slide agglutination tests and standard tube agglutination titres of 1:160 or more. When performed, blood culture was often diagnostic. The children were treated with streptomycin for 2 weeks plus either tetracyclines or trimethoprim-sulphamethoxazole for 3 weeks. Treatment was well tolerated. Relapse was observed in one case. CONCLUSION: Brucellosis nowadays affects children in an occupational pattern. As symptoms, signs and first-line laboratory findings are not characteristic, agglutination tests and blood culture should be performed in any child with prolonged fever. Treatment is effective, but prevention of the disease by animal testing and education of high risk families is indicated.

Abstract: The success of the use of liposomes as drug carriers depends on both their formulation and the method of preparation. We have carried out a series of in vitro studies using different formulations and preparation methods, with the aim of obtaining a type of liposome which is efficient in the treatment of brucellosis. On the basis of results obtained in studies of stability at 37 degrees C in the presence of serum lipoproteins and of the activation of phagocytic cells and antibiotic transport to the interior of monocytes infected with Brucella abortus, we conclude that the most suitable vesicles are positively charged, stable plurilamellar vesicles (phosphatidylcholine, 30% cholesterol, and 10% stearylamine). Gentamicin incorporated into these cationic liposomes completely eliminated all of the intracellular Brucella organisms (4.6 logs), while free gentamicin was capable of reducing the number of intracellular bacteria by only 0.3 log.

Abstract: Seven hundred and sixty-three camels from 400 herds of local Sudanese camel breeds were investigated for the prevalence of mastitis, identification of its bacterial causes and determination of the leucocyte contents of camel's milk. One hundred and forty-nine (19.5 per cent) of the 763 camels examined were diagnosed as mastitis cases based on clinical signs. One hundred and fifty-nine (47.3 per cent) of the 336 randomly selected milk samples were reactive in a rapid mastitis test and 16 of the 153 tested samples contained Brucella abortus agglutinating antibodies. Streptococcus, Staphylococcus, Micrococcus and Aerobacter species and Escherichia coli were found to be the main causes of mastitis (in descending order). The leucocyte contents of the 757 milk samples ranged from < 5 x 10(5) to > 7.5 x 10(6) leucocyte ml-1 and 42.8 per cent of the samples contained < 5 x 10(5) cells ml-1. Neither significant correlation between the leucocyte content of milk and isolated bacterial species nor significant variation in leucocyte contents during different stages of lactation were detected.

Abstract: The possibility of using TV microscopic analysis for the laboratory diagnosis of salmonellosis and brucellosis is considered. The comparative analysis of this method and standard serodiagnostic methods was made in the study of sera taken from 134 salmonellosis patients 69 chronic brucellosis patients. The study demonstrated the advantages of the method of MIA (rapidity, high sensitivity, specificity), making it possible to recommend its wide use for the laboratory diagnosis of salmonellosis and brucellosis.

Abstract: Brucellosis in dairy cattle is endemic in Zimbabwe. The prevalence continues to be monitored intensively. Only milk and serum samples are routinely screened. Attempts to culture Brucella spp. from clinical specimens are seldom made. Consequently, incidence of various Brucella spp. within Zimbabwe is virtually unknown, despite the high serepositivity reported. This information is paramount in understanding the transmission cycle and is also significant to public health; particularly as B melitensis infects humans more often than do the other brucellae. This paper describes the results of bacteriological and serological investigations of brucellosis in a dairy from near Bulawayo. The said farm was selected for the present pilot study because of the high incidence of reported abortion. The milk ring test was employed to test the bulk pooled milk samples once a month for 14 months. The test was recorded highly positive on all 14 occasions. To locate reactors, milk samples from 36 individual cows were similarly tested. Of these, 21 (almost 59%) were found to be reacting positively. One hundred and seventy-seven animals were marked for serotesting. Of these 40 (approximately 25%) showed quite high serum titres (> 1:360) in both the STT and the Rosebengal test. The farmer was advised to havet all abortions full investigated. However, all the clinical material from cases of abortion, except one, were received in an advanced state of putrefaction. From this, Brucella was isolated on culture from stomach contents and cotyledons. The isolates from both the sites were characterized in detail, employing dye inhibition, phagetyping; the oxidative metabolic test and agglutination with monospecific sera. Both the isolates belonged to B. abortus biovar I, which was confirmed by the Central Veterinary Research Laboratory, Weybridge. The significance of isolation and the need to intensify similar studies have been discussed.

Abstract: Reports on simultaneous central and peripheral nervous system involvement in a patient with brucellosis are very rare. We report of one young female patient with a long history of consumption of non-pasteurized dairy products in which clinical and laboratory findings confirmed the existence of an active brucellosis with nervous system impairment. Cerebrospinal fluid (CSF) analyses were negative. Electrophysiology and positive findings on sural nerve biopsy complemented the diagnosis of polyneuroradiculomyeloencephalitis. Treatment with a combination of doxycycline and rifampin for 2 months was successfully applied. No relapse or sequelae occurred in the patient after 12 months of follow up.

Abstract: An indirect enzyme immunoassay for detection of antibody to Brucella abortus in bovine milk was developed and validated using 6238 milk samples from Canadian herds (brucellosis free) and 202 samples from herds infected with B. abortus (from Argentina and Chile). The assay utilized lipopolysaccharide as the antigen, immobilized on the polystyrene matrix, whole milk to test and a mouse monoclonal antibody, specific for an epitope of bovine IgG1, conjugated with horseradish peroxidase. The sensitivity of the assay was 95.2% +/- 3.7% at a confidence limit of 95% for samples from B. abortus infected herds obtained from chile and 98.7% +/- 0.3% at a confidence limit of 95% for samples from similar herds in Argentina. Of the negative milk samples tested, 77 gave a result above the threshold value of 0.200 optical density units. When the 77 false positive samples were retested using 7.5 mM (final concentration) of EDTA and ethyleneglycol-bis-aminoether-N,N,N', N'-tetraacetic acid (EGTA), the number of false positive reactions was reduced to 3, giving a diagnostic specificity of 99.95%. The divalent cation chelating agents did not affect positive reactions and the sensitivity remained the same. Based on control samples included with each assay, the performance of the assay was consistent.

Abstract: BACKGROUND: In spite of the great effort that has been made in recent years in Castilla-La Mancha to control brucelosis, a lack of awareness on the part of producers and consumers leads to major epidemic outbreak, such as the one described below. METHODS: A description of the outbreak is described and a study is conducted of cases and controls to determine the factors which are responsible for the epidemic. Unadjusted and adjusted Odds Ratios (O.R.) are obtained together with their confidence intervals, for the main epidemiological factors studied. RESULTS: A total of 81 cases of brucelosis were recorded in a period of 25 weeks. All the cases occurred in the same borough or were in some way linked to it. In the case and controls study no differences were found with regard to age, sex, contact with livestock or the consumption of pasteurised milk or cheese. A strong link was established between the illness and the consumption of home-made cottage cheese prepared by a small-scale producer in the borough, (O.R. = 311.9; C.I. = 95% = 41.48-12735)., whose livestock turned out to be infected with Brucella Mellitensis. CONCLUSIONS: This outbreak showed that in Spain, there is a risk of contracting brucelosis by consuming non-pasteurised dairy products, particularly cheese, when these are not subjected to the normal sanitary and health controls. The benefits of epidemiological research in the search for cases and determining the factors responsible for the outbreak are also demonstrated. An intensification of controls, the cleansing of the herds and flocks and health education in general, are suitable instruments for controlling this zoonosis in Spain.

Abstract: There is an increasing need for the veterinary profession to meet the new challenges brought about by increased and intensified livestock production. These challenges consist of control and eradication of diseases, the humane treatment of animals and the prevention of transmission of disease from animals or animal products to man. Examples are given of the contribution which regulatory medicine activities have made to the prevention/control/eradication of screw worm, foot-and-mouth disease, Venezuelan equine encephalomyelitis, Newcastle disease, bovine brucellosis, hog cholera, and certain other diseases in North America.

Kadohira M, McDermott JJ, Shoukri MM, Kyule MN. Variations in the prevalence of antibody to brucella infection in cattle by farm, area and district in Kenya. Epidemiology and infection. 1997 Feb; 118(1); 35-41. [PubMed: 9042033].

Abstract: Variations in the sero-prevalence of antibody to brucella infection by cow, farm and area factors were investigated for three contrasting districts in Kenya: Samburu, an arid and pastoral area: Kiambu, a tropical highland area; and Kilifi, a typical tropical coastal area. Cattle were selected by a two-stage cluster sampling procedure and visited once between August 1991 and 1992. Schall's algorithm, a statistical model suitable for multi-level analysis was used. Using this model, older age, free grazing and large herd size (> or = 31) were associated with higher seroprevalence. Also, significant farm-to-farm, area-to-area and district-to-district variations were estimated. The patterns of high risk districts and areas seen were consistent with known animal husbandry and movement risk factors, but the larger than expected farm-to-farm variation within high risk areas and districts could not be explained. Thus, a multi-level method provided additional information beyond conventional analyses of sero-prevalence data.

Abstract: Dynamic changes in patient demography that are currently altering the regional epidemiology of brucellosis attest to the need for physicians to reacquaint themselves with a disease that has been largely forgotten in the United States. This is especially the case in California, which has a large immigrant population and where brucellosis clearly appears to have evolved from an occupational to a food-borne illness. In our recent clinical experiences with several cases of brucellosis, food-borne transmission of the organism is the presumptive cause of the disease, as no causes were associated with occupational risks for exposure to the organism. This suggests that given a clinical history consistent with brucellosis, physicians working with patient groups at risk for food-borne exposure must inquire about the ingestion of unpasteurized dairy products specifically and early during the patient visit. A history of travel to areas endemic for brucellosis may further aid diagnosis. Although a predominance of nonspecific clinical signs and symptoms (such as fevers or arthralgias) often makes the clinical diagnosis difficult, the frequency and characteristic patterns of localized disease should heighten clinicians' index of suspicion and lower the threshold for a serologic investigation. Prominent musculoskeletal complaints (especially back pain) accompanied by constitutional symptoms such as fever, malaise, and weight loss may be consistent with brucellosis and a history of unpasteurized dairy ingestion should be elicited. Radiographic evidence that localizes the source of back pain as caused by sacroiliitis or spondylitis is highly suggestive of brucellosis in appropriate patients. In such cases, serologic tests should be persuaded early if warranted by the clinical impression.

Abstract: BACKGROUND. Brucellosis is considered the most important zoonosis in Jordan with high prevalence among man and livestock. METHODS. This study was carried out on high risk people in 1992 in order to assess the seroprevalence of brucellosis in northern Jordan. The sera of 1236 individuals (636 at high risk and 600 controls) were evaluated using the Rose Bengal plate agglutination test (RBPT) and enzyme-linked immunosorbent assay (ELISA-IgG) tests. RESULTS. A significantly (P < 0.05) higher seroprevalence of brucellosis among high risk people (8.2 percent) compared to the control sample (0.5 percent) was found. The overall seroprevalence was significantly higher among sheep farmers and meat handlers than in other occupations tested. The seroprevalence increased with age and years at work, but was not influenced by sex or locality. The results indicated a higher seroprevalence among veterinarians in northern Jordan, compared to central Jordan but the difference was not significant. Seroprevalence was present only among veterinarians working in clinics especially in the working age group (34-43 years). CONCLUSION. The results of this study emphasized the importance of contact infections, namely contact with infected animals and their products, as a method of transmission of brucellosis compared to ingestion of contaminated animal products.

Abstract: Liposomes of the stable multilamellar type, which previously demonstrated great efficiency in antibiotic transport [Vitas et al: Antimicrob Agents Chemother 1996;40:146-151] were used in this study as carriers of gentamicin for treatment of mice lethally infected with Brucella abortus. Treatment with gentamicin in positively charged liposomes produced a protective effect when it was administered 1 day after lethal challenge (70% of protection). On the other hand, the use of free gentamicin or in liposomes with a negative net charge did not produce a protective effect. Moreover, the results reported here also indicated that cationic liposomes themselves had a therapeutic effect on the course of the infection (up to 50% of protection). In conclusion, we observed that cationic liposomal encapsulation of gentamicin resulted in an enhancement of the therapeutic activity of free gentamicin in this mouse model of acute infection.

Malik GM. A clinical study of brucellosis in adults in the Asir region of southern Saudi Arabia. The American journal of tropical medicine and hygiene. 1997 Apr; 56(4); 375-7. [PubMed: 9158043].

Abstract: One hundred four Saudi patients with brucellosis who were admitted to Abha General Hospital in the Asir region of southern Saudi Arabia were studied. All the patients had Brucella melitensis infection; 61.5% were females while 38.5% were males. Their mean age was 32 years. Most of the patients (61.5%) lived in the lowland (Tihama) and the majority were shepherds (84.6%). The most common presenting symptoms were fever (100%), sweating (96.2%), headache (76.9%), joint pains (76.9%), and backache (73.1%). Physical findings included fever (96.2%), hepatomegaly (46.2%), splenomegaly (42.3%), tenderness over the spine (30.8%), arthritis (26.9%), and lymphadenopathy (19.2%). Mild anemia, leukopenia, and relative lymphocytosis were common. A history of raw milk ingestion was an important factor in disease transmission (84.6%), followed by close animal contact (73%) and raw liver consumption (63.3%). The study shows the effectiveness of several drug combinations in the treatment of brucellosis and the low relapse rate if the treatment is prolonged for not less than six weeks.

Abstract: We report a case that, to the best of our knowledge, is the only published instance of infection of a pacemaker and its leads by Brucella melitensis. Furthermore, this case suggests that B. melitensis may be able to persist around pacemaker devices despite its having been eliminated from the rest of the body. The patient was a sheep shearer who had just undergone a 45-day course of antibiotic therapy for brucella and had been considered cured on the basis of negative blood cultures.

Abstract: A sero-prevalence survey was carried out on 108 workers of Djibouti slaughter-house in order to search out the carriers of agents of zoonotic diseases. The sera revealed prevalences of 6.5% for brucellosis, 0.9% for chlamydiosis and 42.6% for toxoplasmosis whereas no positive reactions were detected for Rift valley fever, Crimean Congo hemorrhagic fever, hydatidosis, and toxocarosis. These data are compared with results obtained from cattle and small ruminants slaughtered in Djibouti.

Abstract: A total of 165 serum samples of abattoir personnel of Delhi were tested by Rose Bengal Plate test (RBPT), Standard tube agglutination test (STAT), Complement fixation test (CFT) and Dot-Enzyme linked immunosorbent assay (Dot-ElISA). The Seroprevalence was 20.60, 12.75, 50.30 and 25.45 percent, respectively on the basis of RBPT, STAT, CFT and Dot-ELISA. Seroprevalence was highest among blood collectors (99.77%) followed by miscellaneous group (72%), animal handlers (68.96%), butchers (68.00%) and sweepers (57.14%). Among veterinarians, Seroprevalance was 28.57%. The Seroprevalence was more among persons of higher age group. The study indicates that abattoir personnel are at high risk to infection with brucellosis.

Abstract: Brucella usually carry two highly homologous genes (omp2a and omp2b) for porin-like proteins. In several B. abortus biovars the omp2a gene has a large deletion compared to other Brucella omp2's. In this study we have measured Omp2 pore activity in planar bilayers. Omp2b exhibits well-defined trimeric channel activity whilst Omp2a forms monomeric pores of variable size which are smaller than Omp2b. No sequence homology exists between Omp2 and porins of known structure, so hydrophobic moment analysis has been used to model their membrane topology. From this it appears likely that the deletion removes the crucial L3 internal loop.

Abstract: A new medium for detecting and enumerating Pseudomonas spp. associated with poultry meat spoilage by a rapid impedance technique was developed, after testing potential growth promoters for eight Pseudomonas strains and inhibitors against eight competing strains (Enterobacteriaceae) able to grow on the medium of Mead and Adams (1977). Four basal media (brain heart infusion, brucella broth, Shaedler broth and Whitley impedance broth (WIB)) and a synthetic medium were evaluated. Whitley impedance broth was the best basal medium for detecting variations in impedance in relation to Pseudomonas growth. The efficiency of WIB was improved by adding compounds which enhanced the growth of Pseudomonas on the synthetic medium. Among the incubation temperatures tested, 22 degrees C proved to be the best compromise between growth of Pseudomonas associated with poultry meat spoilage and inhibition of competitors. Among the 15 inhibitory substances evaluated against Pseudomonas competitors, five were chosen for inclusion in the final medium: metronidazole, carbenicilline, cetrimide, cycloheximide and diamide (MCCCD medium). Preliminary results obtained from experiments with beef and pork meat showed that this medium could also be used without diamide and at an incubation temperature of 25 degrees C. The impedance technique using MCCCD medium was then compared with an official method which uses the medium of Mead and Adams (1977) on 106 samples of poultry neck skin. The linear regression coefficient between the two techniques was approximately r = 0.85. Impedance was able to detect 10(3) Pseudomonas g-1 within less than 19 h making it a promising technique for predicting poultry meat spoilage.

Abstract: Neurobrucellosis accounts for <1% of cases of brucellosis in children. Six new cases of neurobrucellosis are presented and data from 39 previously published cases are analysed. The incidence is equal in males and females, and the source of infection is likely to be unpasteurised milk. Clinical presentation varies from severe meningoencephalitis or peripheral neuropathy/radiculopathy to behavioural disturbance. Diagnostic certainty requires isolation of the organism from the CSF, but as this is rarely possible serological diagnosis can be performed with the Coombs test on the CSF. Treatment requires combination antibiotic therapy and should continue for at least 8 weeks.

Abstract: The Milk Ring, Serum Agglutination, Biological, Mercaptoethanol and Rose Bengal Plate Tests were used in combination to diagnose brucellosis in cows in some herds supplying milk to Johannesburg. The work was done in conjunction with routine testing of bulk milk supplies for public health control. 4 769 cows in more than 76 herds were involved in the tests. Results confirm that no single test can demonstrate all infected cows in a herd and that repeated testing is necessary. In an infected herd in which Strain 19 adult vaccination has been used some negative and suspicious cows will have to be sacrificed in eradicating the disease.

Gudoshnik AN. [Characteristics of the epidemiology of deer brucellosis in the residents of a city located in a deer raising area]. Zhurnal mikrobiologii, epidemiologii, i immunobiologii. 1976 Jun; (6); 134-5. [PubMed: 948953].

Abstract: An experiment was conducted to investigate the effects of the lectin, Concanavalin A (Con A), contained in raw Jack bean (JB) (Canavalia ensiformis, L.) seeds on the immunological response of broilers. A maize-soybean meal basal diet was prepared to which either 2.5, 5, or 10% of ground raw Jack bean (RJB) seeds was added. The RJB seeds contained 24 g Con A/kg on a dry matter basis, as measured by rocket immunoelectrophoresis. Similar diets were prepared by using the same levels of JB after toasting at 190 C for 16 min. In addition, the basal diet was pair-fed to groups of chicks at the level of feed intake of chicks fed the 10% RJB diet. Each diet was fed to six groups of six chicks for 6 wk. At 5 wk, 15 of chicks from each diet were immunized against Brucella abortus (BA) and the anti-BA antibody titers were determined 1 wk later by ELISA. Antibody production against Con A was also measured by the same method. Binding of Con A to intestinal villi and subsequent endocytosis were confirmed by microscopic examination using a specific peroxidase-antiperoxidase-staining technique. Performance was recorded weekly. Feed intake and weight gain were reduced (P < 0.05) only by the diet containing 10% RJB, indicating that broiler chicks can tolerate daily intakes of 100 mg of Con A over 6 wk without affecting growth. Toasted JB diets supported adequate chick performance. The antibody response to BA did not differ with dietary treatment. Serum from chicks fed raw JB also contained antibodies against Con A. The bursa of Fabricius, thymus, spleen, and pancreas dry weights, as a percentage of dry body weight, were not affected by the experimental diets. The data indicated that Con A binds to the cells of the gastrointestinal tract, passes into the general circulation and, eventually, elicits an immunological response without affecting the production of antibodies to BA.

Abstract: Meat production is extremely important for the economy of Argentina, and represents 35%-40% of all national agricultural production. Cattle are distributed throughout the country, with the highest density in the Pampa region, which hosts 66% of the national herd and produces 80% of the total meat production in Argentina. This region has a long tradition of livestock rearing, and the practice of rotation between agriculture and animal husbandry ensures an ecological equilibrium. Meat production in Argentina is based on the exploitation of natural resources, with traditional extensive grazing, which offers advantages when compared with other countries. The composition of meat depends on factors such as breed and age, and on the diet of the animals. The authors demonstrate that meat produced under extensive grazing conditions in Argentina has less intramuscular fat and cholesterol than that produced in intensive systems which have a grain-rich diet. The authors analyse the situation of the country with regard to diseases which can be transmitted by meat and which might affect public health, particularly bovine spongiform encephalopathy (BSE), brucellosis, tuberculosis, salmonellosis, campylobacteriosis, colibacillosis and taeniosis. The fact that Argentina is free from BSE is emphasised, and the prevention programmes which have been implemented to control other diseases are described. In addition, the authors discuss tests which are conducted to detect the presence of residues of chemical contaminants in meat. Emphasis is given to the quality of infrastructure, equipment and operation of the mechanised meat-processing plants, which guarantee the hygienic safety of the products. The authors conclude that meat and meat products from Argentina offer a high level of safety for consumers.

Abstract: Foodborne diseases, in particular those related to meat and meat products, have recently become a matter of great public concern. Sheep and goat meat can transmit infections and diseases either through handling during preparation procedures or as a result of ingestion by the consumer. The authors highlight the second route of contamination in relation to meat and meat products from small ruminants in European countries. Among the most important diseases transmitted by mutton and goat meat, toxoplasmosis remains the greatest threat, particularly in immuno-compromised people and in pregnant women. Other pathogens which may be associated with the consumption of meat from small ruminants include: Clostridium perfringens, Cryptosporidium parvum and Campylobacter jejuni. As with other ruminant species, Escherichia coli O157:H7 can be considered as an emerging pathogen, for which control efforts must be made. The classical zoonoses (brucellosis, Q fever, hydatidosis) are also presented here, although the major source of contamination for these diseases remains contact with infected animals or the handling of carcasses. The fact that the association of foodborne diseases with mutton and goat meat is less frequent than with the meat of other animal species should be noted, for the following reasons: a) lower levels of production; b) less intensive production, leading to a weaker microbial contamination; c) mutton and goat meat are subjected to processing less often than other meats; d) the usual boiling or cooking processes.

Abstract: Milk and dairy products harbour a natural microbial flora and/or other micro-organisms, which vary within the wide range of products available on the French market. The origin of contamination by pathogenic bacteria varies with the type of product and the mode of production and processing. Contamination of milk and dairy products by pathogenic micro-organisms can be of endogenous origin, following excretion from the udder of an infected animal. Contamination may also be of exogenous origin, through direct contact with infected herds or through the environment (e.g. water, personnel). Treatment and processing of milk can inhibit or encourage the multiplication of micro-organisms. The authors describe the relevant aspects of bacterial physiology and ecology, the occurrence of bacteria in dairy products, and the public health significance for each of the principal micro-organisms found in such products. Bacteria most frequently involved are mycobacteria, Brucella sp., Listeria monocytogenes, Staphylococcus aureus and enterobacteria (including toxigenic Escherichia coli and Salmonella). At present, systems of testing and surveillance are required for the control of pathogenic bacteria in milk and dairy products, as specified by regulations currently being developed for all countries in the European Union. Preventive measures should take into account the well-established facts concerning the potential microbiological impact of pathogenic bacteria on milk and dairy products. There should be increased recourse to risk analysis methods to assess the threat to the consumer with regard to the presence of pathogenic bacteria in food.

Abstract: An ambitious programme to eliminate pork as an important source of human salmonellosis was initiated in Denmark in 1993 by the Ministry of Food, Agriculture and Fisheries. The programme comprises control of feedmills, breeding and multiplying herds, slaughter herds and slaughter plants, as well as the final product, fresh pork. As a consequence, the level of occurrence of Salmonella spp. in fresh pork produced in Denmark is approximately 1%. Yersinia enterocolitica 0:3 infections are common in slaughter pig herds in Denmark, and pork is considered to be the only source of human infection in the country. The incidence of pork-related occurrences of human salmonellosis and yersiniosis in 1996 was approximately nine cases per 100,000 inhabitants for both diseases. All swine in Denmark are screened for Trichinella spp. infection, although no positive results have been obtained since 1930. Swine are not considered to be a source for Campylobacter jejuni or enterohaemorrhagic Escherichia coli in Denmark. Listeria monocytogenes can be detected in relatively high rates in pork: however, the incidence of human listeriosis is only 0.5 cases per 100,000 inhabitants. Toxoplasma gondii antibodies have been demonstrated in 3% of slaughter pigs, though the importance of pork as a source of infection is probably very low. Denmark is officially free from Brucella abortus, B. melitensis and Mycobacterium bovis.

Abstract: Wild swine populations (Sus scrofa) are present in many regions of the world. Large feral populations in North America and Australia are principally derived from introduced domestic pigs. In Europe, most wild boar are found in Germany and Poland. While wild swine are certainly a significant reservoir of infection for domestic swine diseases (for example, African swine fever virus in wild boar in Sardinia), these swine generally do not constitute a major public health risk. Brucella suis infections and trichinellosis are the most important zoonotic threats to public health.

Abstract: Two flow cytometric assays are described herein. The single cytometric test (SCT) detects antibodies to either Brucella abortus or Staphylococcus aureus in the serum or milk of a cow or water buffalo. The double cytometric test (DCT) detects both anti-B. abortus and anti-S. aureus antibodies concurrently. In the SCT, the sample to be tested is incubated in succession with the antigen (either B. abortus or S. aureus) and the proper secondary antiserum (fluorescein isothiocyanate-labelled rabbit anti-cow immunoglobulin antiserum or rabbit anti-water buffalo immunoglobulin antiserum). In the DCT, the sample to be tested is incubated first with B. abortus and S. aureus antigens and then with the secondary antiserum. The B. abortus antigen used in the DCT is covalently bound to 3-microm-diameter latex particles. The difference in size between B. abortus and S. aureus permits the establishment of whether the antibodies are directed against one, the other, or both antigens. When compared to the complement fixation test, the SCT and DCT each show a specificity and a sensitivity of 100%. The SCT has been used previously to detect anti-S. aureus antibodies. Here its use is extended to the detection of anti-B. abortus antibodies. The DCT is described here for the first time. The DCT appears to be useful for large-scale brucellosis eradication programs. It offers the possibility of using one test to identify animals that are serologically positive for both B. abortus and S. aureus.

Abstract: OBJECTIVE: To determine the effect of source of cattle and distance cattle were transported to a commercial slaughter facility on prevalence and severity of carcass bruises in mature beef cows. DESIGN: Epidemiologic investigation. SAMPLE POPULATION: Carcasses from 3,955 beef cows from 11 states assembled in 89 procurement lots. PROCEDURE: Each carcass was scored for bruise severity and location. Source of cattle was categorized. Distance from source to slaughter facility was determined. An ANCOVA was used to determine effects of variables on carcass bruises. RESULTS: Mature beef cows marketed through livestock auctions conducting first-point testing for brucellosis, especially when transported longer distances (> 325 km) to slaughter facilities, had a greater number and severity of carcass bruises than cows originating from ranches or livestock auctions not conducting first-point testing. There was an increase in number of rib bruises in cows originating from livestock auctions not conducting first-point testing. Prevalence and severity of bruises were not significantly affected by transportation distance between cows originating from auctions not conducting first-point testing and cows originating from ranches. CLINICAL IMPLICATIONS: A strong association existed between handling for brucellosis testing prior to sale for slaughter and distance transported to slaughter facility with carcass bruises in mature beef cows. Bruises are major quality defects that decrease carcass value and slaughter-cow prices. Repeated handling and restraint as well as long-distance transport are issues to consider regarding the responsibility of the livestock industry to provide for the safety and well-being of cattle sold for slaughter.

Abstract: OBJECTIVE: To compare prevalence estimates of brucellosis (BR) in adult beef cattle that originated from different states and regions of Mexico and that were shipped direct-to-slaughter into Texas during 1995. DESIGN: Epidemiologic survey. ANIMALS: About 65,000 adult beef cattle. PROCEDURE: Blood samples were collected during postmortem examinations and were tested for serum antibodies to Brucella abortus, using the particle concentration fluorescence immunoassay and automated complement-fixation test. Prevalence estimates and 95% confidence intervals of BR were calculated by state of origin in Mexico. The difference among prevalence estimates of BR in cattle from different states and regions was tested for significance (P < 0.05), using the proportion test. RESULTS: On the basis of serologic test results, the overall prevalence estimate of BR was 0.32%. The prevalence estimate of BR in cattle from the state of Chihuahua (0.10%) was significantly different than that in cattle from the states of Nuevo Leon (0.23%), Zacatecas (0.34%), Durango (0.47%), Chiapas (1.81%), Tamaulipas (2.71%), Aguascalientes (7.89%), and Campeche (12.24%). In addition, prevalence estimates of BR in cattle were significantly different among the northern (0.22%), south-central (3.18%), and south coastal (9.42%) regions of Mexico. CLINICAL IMPLICATIONS: Results of this study indicate that the number of cattle exposed to B abortus may be significantly different among states and regions of Mexico. Current import sanitary requirements should continue to mitigate potential risk of transmission of BR from sexually intact cattle of Mexican origin to Texas cattle.

Abstract: Polystyrene plates (96 well) were sensitized with Brucella abortus smooth lipopolysaccharide (SLPS) antigen and then air dried at room temperature (RT) for about 1 hour to dry. Dryness was judged complete when a buffer meniscus was absent from the bottom of the well. The plates were resealed kept on the bench at RT for the duration of the study. Testing was done over 13 months by competitive and indirect ELISAs (C- and IELISA) for bovine antibody to B. abortus using panels of sera from B. abortus S19 vaccinated, unexposed and infected cattle. Testing revealed that consistent results were obtained over the test period suggesting that air drying may be a suitable alterative for storage of plates sensitized with some antigens, in particular, smooth lipopolysaccharide.

Abstract: This article presents an update of several emerging or reemerging pathogens: Yersinia, Cryptosporidia, Cyclospora, Brucella, and Mycobacterium. All of these zoonotic pathogens show evidence of food borne transmission. Yersiniosis is presented as an emerging pathogen that has as its major route of transmission preparation and consumption of pork products. New evidence is presented that supports the transmission of brucellosis via the food chain, especially through contaminated raw milk and cheese. While TB has limited transmission via raw milk, it is highlighted as a reemerging infection due to the development of multiple drug resistance. Public health veterinarians stand in an excellent position to recognize these emerging diseases and apply intervention strategies to prevent and control these infections in the future. This article is intended to raise their consciousness as to the management and medical practices that can diminish food borne transmission.

Abstract: Although several outbreaks of Brucella melitensis infection have been reported among laboratory workers or goat cheese consumers, outbreaks related to rural labour have been rarely studied. An outbreak of human brucellosis among farm workers of Argentina was studied and revealed a close relationship with an epidemic of caprine abortions which occurred shortly before on the same farm. High rates of B. melitensis infection were found among goats. Active brucellosis was diagnosed in 33 subjects (14 with positive blood culture for B. melitensis), while other 27 did not show evidence of illness. While 25 of the brucellosis active patients were rural workers, only 5 of the healthy subjects were engaged in rural labour. Active brucellosis was diagnosed in 91.3% of the subjects in continuous contact with goats and in 32% of those having an occasional contact with the animals. All the 60 subjects denied consumption of goat cheese or milk. As shown here, epidemic human infections by B. melitensis may develop among people frequently in contact with infected goat herds or goat manure.

Abstract: Zoonoses are diseases, the infections of which can be transmitted between man and animals. Only a few are of importance with respect to poultry meat and meat from cattle, sheep, horses and goats. Advances in the control of diseases such as tuberculosis, brucellosis and trichinosis in animals have reduced the hazards posed to workers in the meat industry and to consumers of meat. However, inspection of animals ante- and post-mortem cannot detect all infectious agents present. This applies particularly to bacteria such as Campylobacter, Salmonella, verotoxigenic and other pathogenic Escherichia coli and Yersinia. Protection of meat workers from infection depends upon taking normal hygienic precautions, which also protect the meat from contamination from the workers. Consumers are exposed to a smaller range of zoonoses than meat workers because they encounter only meat that has passed inspection. In addition, heavily contaminated parts of the animal, such as the hide, feathers and viscera have been removed. Further advances in making meat safer are likely to result from the introduction of Integrated Quality Assurance systems. These involve identifying, monitoring and keeping records of the disease status and treatment of each animal (or poultry flock) so that its history is known when it reaches the abattoir. They should also include programmes aimed at minimising colonisation by zoonotic bacteria such as camplyobacters, salmonellas and verotoxigenic Escherichia coli.

Abstract: Conventional methods of ensuring the safety and soundness of cows' milk for human consumption, such as pasteurisation, are not always practical in poor socioeconomic conditions or in rural communities that lack modern amenities. Activation of lactoperoxidase (LP) system and souring of milk were investigated as potential alternative methods to sustain the safety of milk by inhibiting certain microorganisms with known pathogenic potential. The activation of the LP-system inhibited the growth of Staphylococcus aureus and Escherichia coli by the order of 2 log values. The inhibition of Brucella abortus was negligible. The replication of Coxiella burnetti in milk was not disturbed even after 17 h of LP-system activation at 20 degrees C, but the outcome of the LP-system treatment on Mycobacterium bovis could not be determined as the conventional culturing technique used to grow this organism did not allow full recovery. Souring inhibited the growth of S. aureus and E. coli also by the order of 2 log values. From the results obtained in this investigation are concluded that the activation of the LP-system and souring can be used to inhibit the growth of S. aureus and E. coli in cows' milk, thereby increasing its safety.

Abstract: Many diseases are transmitted to man by consumption of contaminated food and drinking water. Orally transmitted diseases are among the main risks for travelers in developing and tropical countries. A variety of clinical manifestations can be observed but the diarrhea is the most common. In many cases bacterial gastroenteritis, typho-paratyphoidal fever, brucellosis, viral hepatitis, and various parasitic diseases can develop after various periods of incubation following consumption of contaminated food or drink with no initial reaction. Vaccination can afford protection against only few diseases. Prevention by applying good hygiene and common sense is the best method. By following the standard list of food precautions, travelers can minimize the main risks. Drug prophylaxis is recommended only under special circumstances. Travel medicine providers must have up-to-date information and possess the persuasive powers necessary to convince travelers to apply recommendations despite the major inconveniences.

Abstract: Brucellosis is an ubiquitous anthropozoonoses in the Republic of Djibouti, but it has only been studied in the City of Djibouti. This retrospective study of 42 cases of human brucellosis diagnosed between April 1993 and April 1995 was carried out at the Peltier Hospital in Djibouti to obtain epidemiological data concerning brucellosis in the Republic of Djibouti. Diagnosis was based on clinical symptoms and positive immunologic tests (Rose Bengal test > 100 UI and Wright serology > 1/80). The following information was obtained for each patient: nationality, ethnic origin, place of residence, age, sex, and risk factors for exposure to infected animals or products. There were 30 men and 12 women including 38 Djiboutis, 3 Somalis, and 1 Ethiopian. Ethnic origin was Afar in 32 cases, Somali in 6 cases, and undetermined in 4 cases. Mean age was 31.6 years. Of the 38 Djiboutis, 15 lived in the district of Djibouti, 17 in the district of Tadjourah, 3 in the district of Obock, and none in districts located in the Southern part of the country. In 18 cases brucellosis involved cattle raisers from the Northern part of the country. In five cases no risk factor could be identified. The incidence of brucellosis in the Republic of Djibouti appears to be comparable to that observed in other highly endemic countries. Further study is needed to determine the exact incidence in man and animals and ascertain the need and feasibility of preventive measures.

Abstract: Broiler breeder hens were fed diets amended with 0 and 10 mg/kg (Trial 1) or 0, 0.2, 1, or 5 mg/kg (Trial 2) of aflatoxin (AF). Fertile eggs collected during 14 d of AF feeding were examined for AF residues. Various immunological endpoints were examined in chicks hatched from these eggs. Eggs collected at 7 d of AF feeding (Trial 1) had 0.15 to 0.48 ng/g of AFB1 and 0.22 to 0.51 ng/g of aflatoxicol, whereas eggs collected at 14 d of AF feeding had 0.05 to 0.60 ng of AFB1/g and 0.19 to 1.20 ng of aflatoxicol/g. In both trials, AF dietary exposure resulted in embryonic mortality and reduction in hatchability compared to controls. The AF progeny chicks in Trial 2 had total anti-SRBC antibodies similar to the controls during the primary antibody response. However, at 5 and 7 d after secondary SRBC injection, the antibody levels in the 1 and 5 mg/kg AF groups were lower than those of controls. Depression in anti-Brucella abortus antibodies occurred only in chicks from the 5 mg/kg AF group. Furthermore, phagocytosis of SRBC and reactive oxygen intermediate production by macrophages from AF progeny chicks were reduced as compared with the control chicks. The findings of this study imply that the progeny chicks from hens consuming a AF-amended diet may be increasingly susceptible to disease owing to suppression of humoral and cellular immunity.

Abstract: BACKGROUND: The appearance of an exceptional number of cases of Brucellosis at the end of 1996 in workers at a slaughterhouse led us to suspect an epidemic outbreak among this group. This study shows the methodology followed in the analysis of this outbreak as well as the results obtained. METHODS: 1.-Epidemiological description of the outbreak: number of animals with brucellosis slaughtered, collection of information from different sources on the number of those affected: from the mutual insurance company, the record of working days lost, an epidemiological monitoring system and a survey amongst the workers. 2.-A case and control study was designed in order to determine, firstly, non-occupational risks--ingestion of fresh cheese or milk and care of animals--and secondly, occupational risks, depending on the job normally undertaken. 3.-To verify this a retrospective cohort study was designed. The exposed group was made up by workers in the slaughter area and the unexposed group comprised the remainder: any worker giving a positive result to the Rose of Bengal test and IgM brucellosis antibodies in serum was considered as a case. RESULTS: The description of the outbreak enabled us to establish that the cases occurred at the moment when most animals were slaughtered, that only occupational risks were relevant, that there were more symptomatic cases than notified ones, and that the slaughter line operators showed higher rates of attack than the remainder of the workers. CONCLUSIONS: This study analyses the possible causal relationship between analyzed exposure and the appearance of cases of brucellosis in workers--a fact which backs the existing scientific evidence on the importance of the respiratory tract as a mechanism of transmission of this disease in the workplace.

Abstract: Since 1990, unusually high rates of false-positive serological reactions (FPSR) in bovine brucellosis screening have been observed in some countries of the European Union. The aim of this survey was to describe this phenomenon in a highly affected French Department, and to evaluate the links between some individual or herd factors and the occurrence of these FPSR. Before 1990, low backgrounds of FPSR were recorded (individual prevalence rate: less than 0.5 per 10,000). The phenomenon burst during the 1990-91 screening campaign, reached a peak in 1992-93 (50.5 per 10,000), and then decreased until the last studied campaign, 1995-96 (9.1 per 10,000). The phenomenon was transient and sporadic within a herd. At the herd-screening level, four assumed risk factors were isolated: (i) the probability of a herd-screening to be positive was closely and positively linked with the herd screening size; (ii) during a given screening campaign, the prevalence of FPSR decreased from December to November; (iii) the presence of at least one goat on the premises increased the risk for the 1992-93 and 1993-94 screening campaigns; and (iv) a previous FPSR in a given herd appeared to be a weak but significant risk factor. At the individual-animal level, herd size, sex and breed did not seem to be linked with FPSR appearance, while young animals were significantly more affected than older ones. However, global variations in herd or individual prevalences remained unexplained. The lack of link between FPSR and brucellosis is strengthened. The hypothesis of a widely spread causal agent with a low individual host susceptibility and/or a low probability of detecting FPSR animals can be supported by these results.

Abstract: The authors intend to elaborate a brucellosis eradication program which could be used in the Amazonian region, an area characterised by extremely extensive animal husbandry practices. Under such conditions, eradication by selection is the only feasible approach. Brucellosis has been successfully eradicated from a herd with 22.1% prevalence of infection by two serological surveys using an indirect ELISA and the complement fixation test.

Abstract: Twenty free-ranging guanaco (Lama guanicoe) in Chubut Province, Argentina, were immobilized for health evaluations. All but two animals appeared to be in good condition. Hematology, serum chemistry, and vitamin and mineral levels were measured, and feces were evaluated for parasites. Serology tests included bluetongue, brucellosis, bovine respiratory syncitial virus, bovine viral diarrhea/mucosal disease, equine herpesvirus 1, infectious bovine rhinotracheitis, Johne's disease (Mycobacterium paratuberculosis), foot and mouth disease, leptospirosis (17 serovars), parainfluenza-3, and vesicular stomatitis. Blood samples from 20 domestic sheep (Ovis aries) maintained in the same reserve with the guanaco were also collected at the same time for serology tests. No guanaco had positive serologic tests. Sheep were found to have antibody titers to bovine respiratory syncytial virus, Johne's disease, leptospirosis, and parainfluenza-3. There was no apparent difference in external appearance or condition, or statistical difference in blood test values, between the animals that were positive or negative for parasite ova.

Falade S. A comparison of three serological tests in the diagnosis of caprine brucellosis. Research in veterinary science. 1978 May; 24(3); 376-7. [PubMed: 97740].

Abstract: The serum agglutination test (SAT), the Rose Bengal plate test (RBPT) and the milk ring test (MRT) were used in the diagnosis of caprine brucellosis. There was a close correlation between the SAT and RBPT when both tests were negative but the RBPT failed to detect 79.82 per cent of sera in excess of 50 iu. Also, owing to the relatively poor milking potential of the Nigerian goat and the false positive results with the MRT, it is concluded that the SAT offers a better serological diagnostic tool for caprine brucellosis in this locality.

Abstract: As the result of the prospective examination of 863 pregnant women in urban and rural consultation clinics for women in Daghestan, a high proportion of them were found to be infected with toxoplasmosis (25.5%), brucellosis (1.85%) and listeriosis (12.2%). The data on the contamination of 1325 women with aggravated obstetric history were confidently higher, constituting 52.0%, 3.3% and 22.2% respectively. The results of the examination of women working on live-stock farms (226 women) and poultry farms (106 women) demonstrated a significantly high frequency of contamination with the above-mentioned zoonotic infections. The data thus obtained were indicative of the necessity of organizing epidemiological surveillance on these infections; for their diagnostics a complex of laboratory methods could be used, though the effectiveness of these methods was different in different nosological forms.

Abstract: An indirect enzyme-linked immunosorbent assay (ELISA) for Brucella abortus antibodies detection in bovine milk and serum samples was validated. The assay use B. abortus smooth lipopolysaccharide as antigen, immobilized on a polystyrene matrix; milk diluted 1:2 or serum diluted 1:50, in a buffer containing divalent cation chelating agents EDTA and EGTA (ethyleneglycol-bis-aminoether-N,N,N',N'-tetraacetic acid) to reduce non-specific reactions; and a mouse monoclonal antibody specific for an epitope of bovine IgG1, conjugated with horseradish peroxidase. A total of 2646 sera and 2119 milk samples from cows older than 24 months were obtained from 12 brucellosis-free herds for at least the previous 5 years. Milk samples were obtained in parallel with serum samples. The remaining 527 serum samples were from dry cows. All cattle were vaccinated with B. abortus strain 19 between 3-10 months of age. Five hundred and fifty-two milk samples and 562 serum samples were obtained from 6 infected herds with abortions where B. abortus was isolated at least once no more than 6 months before sampling. The complement-fixation test (CFT) on serum samples was considered the gold standard. Serum samples were also tested with the official screening test: the buffered plate antigen (BPA) test. The cut-off point was determined using receiver-operating characteristic (ROC) analysis. For milk samples, it was fixed at 36 percent positivity (PP) giving a sensitivity of 99.6% with a 95% confidence interval (CI) of 98.6-99.9%. The specificity was 99.1% (CI 98.9-99.4%). For serum samples, the cut-off was fixed at 53 PP giving a sensitivity of 99.6% (CI 98.6-99.9%) and a specificity 98.6% (CI 98-99%). The BPA test showed a relative sensitivity of 99.6% (CI 98.6-99.9%) and a relative specificity of 98.6% (CI 98.1-99%). Our results indicate that the indirect ELISA is a highly sensitive and specific test and can be adapted to process a large number of samples.

Abstract: To identify contributing factors for cheese-associated outbreaks, we reviewed all cheese-associated outbreaks of human illness reported to the Centers for Disease Control and Prevention (CDC) with onsets during 1973 to 1992. The infrequency of large, cheese-associated outbreaks was notable because such outbreaks had been a frequent public health problem before the mid-20th century. Of 32 reported cheese-associated outbreaks, 11 attributed to manufacturing errors caused most of the illnesses and hospitalizations and all 58 deaths. Important factors in these 11 outbreaks were manufacturing cheese with raw or improperly pasteurized milk and postpasteurization contamination. If current Food and Drug Administration sanitary requirements for cheesemaking had been met, these outbreaks would have been preventable. In two outbreaks of Salmonella infections, fewer than 10 Salmonella per 100 g of cheese were detected. In two outbreaks of Brucella infections, efforts to recover the pathogen from the implicated cheese were unsuccessful, emphasizing the inadequacy of end product testing for assuring consumer safety. Curing cheeses kills most bacteria present in cheeses; however, evidence from sources other than the CDC Foodborne Disease Outbreak Surveillance System suggests that curing alone may not be a sufficient pathogen control step to eliminate Salmonella, Listeria, and E. coli O157:H7 from cheese.

Abstract: An indirect ELISA (iELISA) using a B. abortus smooth lipopolysaccharide (S-LPS) as coating antigen and a polyclonal anti-sheep IgG peroxydase-labeled serum as conjugate was developed. The iELISA was assessed in comparison to the Food and Agriculture Organisation/International Atomic Energy Agency (FAO/IAEA) standard bovine ELISA kit (IAEA kit) using an anti-bovine IgG conjugate that cross-reacts with sheep antibodies, and with complement fixation test (CFT) and Rose Bengal Plate Test (RBPT). The evaluation was performed on sera from ewes experimentally infected with Brucella melitensis 53H38 (H38), using negative and positive sheep reference sera. No significant difference was found as regards to the specificity, the lower limit of detection and the estimated sensitivity of the two iELISAs. This suggests that an anti-bovine conjugate could allow the development of only one ELISA protocol for all ruminants. The iELISA, if well standardized, proved to be a good screening test able to be used either alone or in addition to RBPT.

Abstract: The transfer of chronically protein-deficient mice to an optimal diet a few days before immunization with sheep erythrocytes, tetanus toxoid or Brucella abortus vaccine either failed to increase antibody production above the level produced by deficient mice, or suppressed the responses to below those produced by deficient mice or normally-fed controls. Transfer to high protein diet on the day of immunization or feeding deficient mice the normal diet for just 2 days at the time of injection produced higher titres than did transfer a few days before immunization. Secondary responses to TT were affected by transfer to the normal diet at priming, rather than at rechallenge. Some mechanisms which may explain these findings and their implications for immunization schedules in malnourished humans are discussed.

Collin JC. [The role of the different immunoglobulins in the milk ring test used for the detection of brucellosis (author's transl)]. Annales de microbiologie. 1976 Aug-Sep; 127B(2); 177-87. [PubMed: 999126].

Abstract: The serum immunoglobulins obtained from one heifer vaccinated against brucellosis, and milk immunoglobulins from a cow suffering from a natural udder infection, were purified. The preparations were more than 99.5 per cent pure. The role of each immunoglobulin in the milk ring test was determined. Fractions sIgA and IgM induce the formation of a broad coloured ring after 1 hour's incubation at 37 degrees C. Under these same conditions, no ring was observed with IgG1 or IgG2 but the coloured agglutinated bacteria were visible at the bottom of the tube. The maintenance of the tubes at 4 degrees for 18 hours increased the sensitivity of the milk ring test, with all immunoglobulins. When the four immunoglobulins were simultaneously present in milk, the formation of the coloured ring prevented the formation of a deposit of agglutinated bacteria on the bottom of the tube. In this case, agglutination at the bottom of the tube occured only at dilutions above that giving the coloured ring. Addition of IgG1 and IgG2 to the mild containing sIgA or IgM slightly increased the title of the milk ring test: one part of the deposit of agglutinated bacteria rose to the surface with the fat globules.